U.S. patent number 5,532,359 [Application Number 08/328,595] was granted by the patent office on 1996-07-02 for ras farnesyl transferase inhibitors.
This patent grant is currently assigned to Board of Regents, The University of Texas System, Genentech, Inc.. Invention is credited to Michael S. Brown, Craig W. Crowley, Joseph L. Goldstein, Guy L. James, James C. Marsters, Jr., Robert S. McDowell, David Oare, Thomas E. Rawson, Mark Reynolds, Todd C. Somers.
United States Patent |
5,532,359 |
Marsters, Jr. , et
al. |
July 2, 1996 |
Ras farnesyl transferase inhibitors
Abstract
Benzodiazepine derivatives are disclosed that act as potent
inhibitors of ras farnesyl:protein transferase. Pharmaceutical
compositions containing these benzodiazepines are provided for
treatment of diseases for which inhibition of the ras
farnesyl:protein transferase is indicated. Also disclosed are
benzazepines of the following general formula (II) having similar
utility as the aforementioned benzodiazepines: ##STR1##
Inventors: |
Marsters, Jr.; James C.
(Oakland, CA), Brown; Michael S. (Dallas, TX), Crowley;
Craig W. (Portola Valley, CA), Goldstein; Joseph L.
(Dallas, TX), James; Guy L. (Dallas, TX), McDowell;
Robert S. (San Francisco, CA), Oare; David (Belmont,
CA), Rawson; Thomas E. (Mountain View, CA), Reynolds;
Mark (So. San Francisco, CA), Somers; Todd C. (Montara,
CA) |
Assignee: |
Genentech, Inc. (South San
Francisco, CA)
Board of Regents, The University of Texas System (Austin,
TX)
|
Family
ID: |
26741701 |
Appl.
No.: |
08/328,595 |
Filed: |
October 25, 1994 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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82202 |
Jun 24, 1993 |
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61961 |
May 14, 1993 |
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Current U.S.
Class: |
540/522; 540/499;
540/523; 540/497; 540/504; 540/558; 540/509; 540/498; 540/493 |
Current CPC
Class: |
C07D
223/16 (20130101); C07D 403/14 (20130101); C07K
5/0606 (20130101); C07K 5/06139 (20130101); C07D
403/12 (20130101); C07D 243/10 (20130101); C07K
5/0821 (20130101); C07K 5/1024 (20130101); C07D
401/14 (20130101); C07K 5/06191 (20130101); C07D
409/14 (20130101); C07D 403/10 (20130101); C07K
5/1027 (20130101); C07K 5/0827 (20130101); C07K
5/0207 (20130101); C07K 5/1013 (20130101); C07D
401/06 (20130101); C07D 405/06 (20130101); C07D
403/06 (20130101); C07D 487/04 (20130101); C07K
5/081 (20130101); A61P 31/04 (20180101); A61P
35/00 (20180101); C07D 243/24 (20130101); C07D
513/04 (20130101); A61K 38/00 (20130101) |
Current International
Class: |
C07D
498/00 (20060101); C07K 5/02 (20060101); C07D
498/04 (20060101); C07K 5/06 (20060101); C07K
5/00 (20060101); C07D 405/00 (20060101); C07D
405/06 (20060101); C07D 243/24 (20060101); C07D
403/12 (20060101); C07D 403/14 (20060101); C07D
403/06 (20060101); C07D 243/10 (20060101); C07D
243/00 (20060101); C07D 403/10 (20060101); C07D
409/00 (20060101); C07D 513/04 (20060101); C07D
409/14 (20060101); C07D 513/00 (20060101); C07K
5/078 (20060101); C07D 223/00 (20060101); C07D
223/16 (20060101); C07K 5/103 (20060101); C07K
5/083 (20060101); C07K 5/08 (20060101); C07K
5/10 (20060101); C07K 5/097 (20060101); C07K
5/117 (20060101); C07D 401/14 (20060101); C07D
401/06 (20060101); C07D 403/00 (20060101); C07D
401/00 (20060101); C07D 487/04 (20060101); C07D
487/00 (20060101); A61K 38/00 (20060101); A61K
031/55 (); C07D 223/16 (); C07D 223/18 () |
Field of
Search: |
;540/522,523
;514/213,217 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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0166357 |
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Jan 1986 |
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EP |
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0167919 |
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Jan 1986 |
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EP |
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0322779 |
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Jul 1989 |
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EP |
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WO92/01683 |
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Feb 1992 |
|
WO |
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WO94/04561 |
|
Mar 1994 |
|
WO |
|
Other References
James, G. et al., "Benzodiazepine peptidomimetics: potent
inhibitors of ras farnesylation in animal cells" Science
260:1937-1941 (1993)..
|
Primary Examiner: Datlow; Philip I.
Attorney, Agent or Firm: Winter; Daryl B.
Parent Case Text
This is a continuation of application Ser. No. 08/082,202 filed on
24 Jun. 1993, now abandoned which application is a
continuation-in-part of Ser. No. 08/061,961 filed on 14 May 1993
now abandoned.
Claims
What is claimed is:
1. A compound represented by structural formula (II): ##STR3015##
where R and R' are independently selected from the group
hydrogen,
halo(F, Cl, Br, I),
C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl,
C.sub.1 -C.sub.6 alkoxy,
hydroxy,
hydroxy-C.sub.1 -C.sub.6 alkyl,
C.sub.1 -C.sub.6 alkylcarbonyl, and
C.sub.1 -C.sub.6 alkyloxycarbonyl;
R.sup.1 and R.sup.2 are independently selected from the group
hydrogen,
C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)-C.sub.1 -C.sub.6 alkyl, and ##STR3016## R.sup.1
and R.sup.2 taken together may form a covalent bond or fused
benzene substituted with R and R';
R.sup.4 and R.sup.4' are independently selected from the group
hydrogen,
halo(F, Cl, Br, I),
C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl,
phenyl, and
benzyl;
R.sup.7 selected from the group
hydrogen,
halo(F, Cl, Br, I),
C.sub.1 -C.sub.6 alkyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl;
W is selected from the group
C(.dbd.O)--NR.sup.7 'R.sup.8,
CH.sub.2 --C(.dbd.O)--NR.sup.7 'R.sup.8,
CR.sup.8' (OH)--CHR.sup.7 R.sup.8,
CHR.sup.8' --CHR.sup.7 R.sup.8,
CHR.sup.8 '--CHR.sup.7 R.sup.8,
CR.sup.8' CR.sup.7 R.sup.8 (E or Z),
C(.dbd.O)--CHR.sup.7 R.sup.8,
CHR.sup.8' --NR.sup.7' R.sup.8,
CHR.sup.8' --O--R.sup.8,
CHR.sup.8' --S(O).sub.u --R.sup.8 where u is 0, 1, or 2,
CR.sup.8' .dbd.N--R.sup.8,
CHR.sup.8' --R.sup.8,
W',
C.sub.1 -C.sub.3 alkyl-W',
C.sub.6 -C.sub.12 aryl-W',
C.sub.6 -C.sub.12 aryl-C.sub.1 -C.sub.3 alkyl-W',
heterocycle-W',
heterocycle-C.sub.1 -C.sub.3 alkyl-W',
C.sub.1 -C.sub.2 alkyl-C.sub.6 -C.sub.10 aryl-W', and
C.sub.1 -C.sub.2 alkyl-heterocycle-W',
where any heterocycle is a 5- or 6-member saturated or unsaturated
ring containing 1 to 3 heteroatoms selected from O, N, and S;
W' is selected from one to three substituents selected from the
group
hydrogen,
SR.sup.9,
SSR.sup.9,
SC(.dbd.O)--R.sup.9,
OR.sup.9,
C(.dbd.NH)--NH.sub.2,
N.dbd.CH--NH.sub.2,
NH--CH.dbd.NH,
R.sup.8, and
V;
R.sup.7' is selected from the group
hydrogen,
benzyl,
C.sub.1 -C.sub.4 alkyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.4 alkyl;
R.sup.8' is selected from the group
hydrogen,
C.sub.1 -C.sub.4 alkyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.4 alkyl;
R.sup.7' and R.sup.8 together with the nitrogen to which they are
bonded may form a pyrrolidinyl or piperidyl ring optionally
substituted with one or two groups selected from
SR.sup.9,
SSR.sup.9,
SC(.dbd.O)--R.sup.9,
OR.sup.9,
C(.dbd.O)NHOH,
NHR.sup.9,
C(.dbd.O)NR.sup.27 R.sup.28, and
V;
R.sup.8 is selected from the group unsubstituted and
substituted
C.sub.1 -C.sub.8 alkyl,
C.sub.1 -C.sub.4 alkyl-Z-C.sub.1 -C.sub.4 alkyl, where Z is S or
O,
C.sub.2 -C.sub.4 alkyl-NR-C.sub.2 -C.sub.4 alkyl,
C.sub.2 -C.sub.8 alkenyl,
C.sub.6 -C.sub.12 arylC.sub.1 -C.sub.3 alkyl,
indol-3-yl-C.sub.1 -C.sub.3 alkyl, and
imidazol-4-yl-C.sub.1 -C.sub.3 alkyl,
where any aryl moiety is optionally substituted with --OR.sup.9 and
V, and
where any alkyl or alkenyl group is optionally substituted with one
to three groups selected from
SR.sup.9,
SSR.sup.9,
SC(.dbd.O)--R.sup.9,
OR.sup.9,
C(.dbd.NH)--NH.sub.2,
N.dbd.CH--NH.sub.2,
NH--CH.dbd.NH,
NH--C(.dbd.NH)--NH.sub.2,
C(.dbd.O)NHOH,
NHR.sup.9,
C(.dbd.O)NR.sup.27 R.sup.28, and
V;
V is selected from the group
COR.sup.10,
SO.sub.3 R.sup.13,
NHSO.sub.2 CF.sub.3,
PO(OR.sup.13).sub.2,
SO.sub.2 NHR.sup.10,
CONHOR.sup.13,
C(OH)R.sup.10 PO(OR.sup.13).sub.2,
CN,
SO.sub.2 NH-heteroaryl where the heteroaryl is a 5- or 6-member
aromatic ring containing 1 to 3 heteroatoms selected from O, N, and
S and where the heteroaryl is unsubstituted or substituted with one
or two substituents selected from the group
OH,
SH,
C.sub.1 -C.sub.4 alkyl,
C.sub.1 -C.sub.4 alkoxy,
CF.sub.3,
halo(F, Cl, Br, I),
NO.sub.2,
COOH,
COO--(C.sub.1 -C.sub.4 alkyl),
NH.sub.2,
NH(C.sub.1 -C.sub.4 alkyl), and
N(C.sub.1 -C.sub.4 alkyl).sub.2,
CONHSO.sub.2 R.sup.15,
SO.sub.2 NHCOR.sup.15,
CONHSO.sub.2 R.sup.13,
CH.sub.2 CONHSO.sub.2 R.sup.15,
NHCONHSO.sub.2 R.sup.15,
NHSO.sub.2 NHCOR.sup.15,
CONHNHSO.sub.2 CF.sub.3,
CON(OH)R.sup.13,
CONHCOCF.sub.3,
CONHSO.sub.2 R.sup.10,
CONHSO.sub.2 R.sup.11,
CONHSO.sub.2 R.sup.13, ##STR3017## R.sup.9 is selected from the
group hydrogen,
methyl,
ethyl,
isopropyl,
phenyl, and
benzyl;
R.sup.10 is selected from the group
hydroxy,
C.sub.1 -C.sub.8 -alkoxy,
C.sub.3 -C.sub.12 -alkenoxy,
C.sub.6 -C.sub.12 -aryloxy,
C.sub.1 -C.sub.6 -alkyl-C.sub.6 -C.sub.12 -aryloxy,
di-C.sub.1 -C.sub.8 -alkylamino-C.sub.1 -C.sub.8 -alkoxy,
alkanoylamino-C.sub.1 -C.sub.8 -alkoxy selected from the group
acetylaminoethoxy,
nicotinoylaminoethoxy, and
succinamidoethoxy, and
C.sub.1 -C.sub.8 -alkanoyloxy-C.sub.1 -C.sub.8 -alkoxy,
C.sub.6 -C.sub.12 -aryl-C.sub.1 -C.sub.8 -alkoxy where the aryl
group is unsubstituted or substituted with one to three of the
groups
nitro,
halo(F, Cl, Br, I),
C.sub.1 -C.sub.4 -alkoxy, and
amino,
hydroxy-C.sub.2 -C.sub.8 -alkoxy,
dihydroxy-C.sub.3 -C.sub.8 -alkoxy, and
NR.sup.11 R.sup.12 ;
R.sup.11 and R.sup.12 are independently selected from the group
hydrogen,
C.sub.1 -C.sub.6 alkyl,
C.sub.2 -C.sub.6 alkanoyl,
C.sub.1 -C.sub.6 alkanoyl substituted with from one to three groups
selected from
nitro,
halo(F, Cl, Br, I),
C.sub.1 -C.sub.4 -alkoxy, and
amino, and
C.sub.6 -C.sub.12 -aryl-C.sub.1 -C.sub.8 -alkyl where the aryl
group is unsubstituted or substituted with one to three of the
groups selected from
nitro,
halo(F, Cl, Br, I), and
C.sub.1 -C.sub.4 -alkoxy;
R.sup.13 is selected from the group
hydrogen,
C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)-C.sub.1 -C.sub.6 alkyl,
phenyl,
benzyl, and
CH.sub.2 --O--COCH.sub.3 ;
R.sup.15 is selected from the group
C.sub.6 -C.sub.14 aryl, heteroaryl, where the heteroaryl is a 5- or
6-member aromatic ring containing 1 to 3 heteroatoms selected from
O, N, and S and where the heteroaryl is unsubstituted or
substituted with one or two substituents selected from the
group
OH,
SH,
C.sub.1 -C.sub.4 alkyl,
C.sub.1 -C.sub.4 alkoxy,
CF.sub.3,
halo(F, Cl, Br, I),
NO.sub.2,
COOH,
COO--(C.sub.1 -C.sub.4 alkyl),
NH.sub.2,
NH(C.sub.1 -C.sub.4 alkyl), and
N(C.sub.1 -C.sub.4 alkyl).sub.2,
C.sub.3 -C.sub.7 -cycloalkyl,
C.sub.1 -C.sub.4 -alkyl, unsubstituted or substituted with a
substituent selected from the group
C.sub.6 -C.sub.14 aryl,
heteroaryl as defined above,
OH,
SH,
C.sub.1 -C.sub.4 -alkyl,
C.sub.1 -C.sub.4 -alkoxy,
C.sub.1 -C.sub.4 -alkylthio,
CF.sub.3,
halo(F, Cl, Br, I),
NO.sub.2,
CO.sub.2 H,
CO.sub.2 --(C.sub.1 -C.sub.4)-alkyl,
NH.sub.2,
N[(C.sub.1 -C.sub.4)-alkyl].sub.2,
NH[(C.sub.1 -C.sub.4)-alkyl],
PO.sub.3 H, and
PO(OH)(C.sub.1 -C.sub.4)-alkoxy, and
(C.sub.1 -C.sub.4)-perfluoroalkyl;
R.sup.16 is selected from the group
CN,
NO.sub.2,
COOR.sup.13,
C.sub.1 -C.sub.6 -perfluoroalkyl, and
CF.sub.3 ;
R.sup.19 is selected from the group
hydrogen,
C.sub.1 -C.sub.6 alkyl,
C.sub.2 -C.sub.6 alkenyl,
C.sub.1 -C.sub.6 alkoxy,
C.sub.2 -C.sub.6 alkoxyalkyl,
CH.sub.2 --O--COCH.sub.3, and
benzyl, where the phenyl moiety is unsubstituted or substituted
with a group selected from
NO.sub.2,
NH.sub.2,
OH, and
OCH.sub.3 ;
X is selected from the group
NR.sup.24 --C(.dbd.O)--R.sup.25,
NR.sup.24 --CH(OH)--R.sup.25, and
NR.sup.24 --S(O).sub.u --R.sup.25 where u is 0, 1, or 2,
R.sup.24 is selected from the group
C.sub.1 -C.sub.6 alkyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl;
R.sup.25 is selected from R.sup.25', ##STR3018## R.sup.25' is
selected from the group C.sub.1 -C.sub.6 alkyl,
C.sub.2 -C.sub.6 alkenyl,
C.sub.1 -C.sub.6 alkylamine,
C.sub.2 -C.sub.6 alkenylamine, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl
where any alkyl or alkenyl moiety is substituted with NR.sup.27
R.sup.28 and one or more groups selected from
SH and
SSR.sup.26 ;
R.sup.26 is selected from
hydrogen,
C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl, and
C.sub.1 -C.sub.6 alkanoyl;
R.sup.27 and R.sup.28 are independently selected from the group
hydrogen,
C.sub.1 -C.sub.6 alkyl,
phenyl,
napthyl,
benzyl,
CH.sub.2 napthyl (a or b),
C.sub.1 -C.sub.6 alkanoyl,
C.sub.1 -C.sub.6 cycloalkanoyl.
C.sub.6 -C.sub.10 aroyl,
C.sub.6 -C.sub.10 arylC.sub.1 -C.sub.6 alkanoyl,
C.sub.1 -C.sub.6 alkylsulfonyl,
C.sub.6 -C.sub.10 arylsulfonyl,
C.sub.6 -C.sub.10 arylC.sub.1 -C.sub.6 alkylcarbamoyl,
cinnamoyl,
heterocyclecarbonyl,
C.sub.1 -C.sub.6 alkoxycarbonyl,
C.sub.6 -C.sub.10 aryloxycarbonyl,
C.sub.6 -C.sub.10 arylC.sub.1 -C.sub.6 alkoxycarbonyl, and
pyroglutamyl;
R.sup.27 and R.sup.28 together with the nitrogen atom to which they
are bonded may form a cyclic amine represented by ##STR3019## or a
cyclic imide represented by ##STR3020## G is selected from
--CH.sub.2 --, O, S(O).sub.u where u is 0, 1, or 2, and NR.sup.28
;
J-M is selected from C.sub.2 -C.sub.4 alkylene and C.sub.2 -C.sub.4
alkenylene;
R.sup.29 is selected from hydrogen and C.sub.1 -C.sub.3 alkyl;
and
pharmaceutically acceptable salts thereof.
2. The compound of claim 1 where R.sup.24 is methyl.
3. The compound of claim 2 that is substantially free of its
diastereomer.
4. The compound of claim 3 that has the R configuration about the
3-carbon of the seven-member ring.
5. The compound of claim 3 that has the S configuration about the
3-carbon of the seven-member ring.
6. The compound of claim 3 wherein
R.sup.1 is phenyl and R.sup.2 is hydrogen.
7. The compound of claim 6 wherein
X is NR.sup.24 --C(.dbd.O)R.sup.25.
8. The compound of claim 7 wherein
R.sup.25 is C.sub.1 -C.sub.6 alkyl substituted with --SSR.sup.26
and NR.sup.27 R.sup.28.
9. The compound of claim 8 wherein
R.sup.26 is C.sub.1 -C.sub.6 alkyl and
R.sup.27 and R.sup.28 are both hydrogen.
10. The compound of claim 9 wherein
--C(.dbd.O)R.sup.25 is the cysteine residue.
Description
FIELD OF THE INVENTION
This invention relates generally to non-peptidyl inhibitors of
farnesyl:protein transferase, an enzyme capable of catalizing
farnesylation of p21.sup.ras and related low molecular weight
G-proteins. More specifically, the instant inhibitors are analogs
of benzodiazepine and structurally related 6-7 fused ring systems.
The invention further relates to use of these inhibitors in
situations where inhibition of posttranslational farnesylation of
p21.sup.ras and related proteins is indicated.
BACKGROUND OF THE INVENTION
Proteins encoded by the ras proto-oncogene act as molecular
switches responding to growth stimuli and signaling to the
intracellular machinery the occurrence of an extracellular event
such as binding of a growth hormone to a growth hormone receptor
molecule. Binding of the hormone to its receptor (the external
signal) switches the ras protein to the "on" position characterized
by exchange of ras bound GDP for GTP. The tightly bound GTP in turn
stimulates downstream target proteins, ultimately triggering a
cascade of reactions leading to specific gene transcription and
ultimately cell division [Barbacid, M., Ann. Rev. Biochem. 56:779
(1987), McCormick F., Nature 363:15-16 (1993)]. The normal (i.e.
non-transformed) ras protein eventually switches to the off
position by hydrolyzing bound GTP to GDP and the cell is poised to
receive the next external signal.
Mutations to the ras proto-oncogene translate into amino acid
substitutions in the GTP binding domain, activating the ras protein
(p21.sup.ras) and biasing this molecular switch in the "on"
position. Thus, the ras transformed cell behaves like a cell with a
faulty switch, signaling extracellular hormone binding when none is
present. Cells transformed in this way grow and differentiate in an
abnormal way.
Transforming ras genes are the oncogenes most frequently identified
in human cancers. Clinical investigations have identified activated
ras genes in a wide variety of human neoplasms, including
carcinomas, sarcomas, leukemias, and lymphomas. It is estimated
that 40% of all human colon cancers and 95% of human pancreatic
cancers contain activated ras oncogenes [Kuzumaki, N. Anticancer
Res., 11:313-320 (1991)].
Recently, it has been discovered that the ras protein must be
properly posttranslationally modified before it can function as a
molecular switch. Stable modification of the carboxy terminus of
ras proteins appears to be essential for correct localization
within the cell membrane so that extracellular signals for cell
growth and differentiation can be correctly passed along to the
intracellular messengers. The ras proteins are posttranslationally
modified by farnesylation of a cysteine residue located four
residues from the carboxy terminus, followed by proteolytic
cleavage of the three following amino acid residues and methylation
of the free cysteine carboxyl. The farnesylation reaction is
catalyzed by a 94 kda heterodimeric Zn.sup.2+ metalloenzyme,
farnesyl:protein transferase, which transfers the farnesyl group, a
15 carbon isoprenoid lipid derived from mevalonate (a cholesterol
precursor), from farnesyl pyrophosphate to the carboxy terminus
cysteine sulfur of ras forming a stable thioether linkage. The
farnesyl:protein transferase recognizes the ras carboxy terminus
consensus sequence, CAAX, where the cysteine (C) is followed by two
aliphatic (A) amino acids (usually valine, leucine, or isoleucine)
and any amino acid X (including methionine). This consensus
sequence or motif is frequently referred to as the "CAAX box" and
is found in other ras related GTP-binding proteins such as fungal
mating factors, nuclear lamins, the gamma subunit of transducin,
rhodopsin kinase, and the alpha subunit of
cGMP-phosphodiesterase.
Surprisingly, this enzyme does not require intact ras protein for
transferase activity and can utilize tetrapeptides with the CAAX
motif as substrates (Reiss et al., Cell, 62: 81-88 (1990)). This
observation suggested that small tetrapeptides like CAAX or
nonpeptide analogs thereof could compete with p21.sup.ras for the
active site of the transferase and therefore might be of
therapeutic utility.
Previously, it had been observed that mutation of the cysteine in
the CAAX carboxy sequence of p21.sup.ras to serine prevented
farnesylation, proteolysis, and methylation (Hancock, J. et al.,
Cell 57:1167-1177 (1989); Reiss et al., PNAS, 88:732-736 (1991)).
Additionally, cells incubated with an inhibitor of mevalonate
synthesis prevented ras farnesylation and the cells were no longer
capable of cell division (Schafer et al., Science, 245: 379-385
(1989)).
These results, taken together, suggest that inhibition of
farnesyl:protein transferase with peptides containing the CAAX
motif would prevent farnesylation of p21.sup.ras and block the
ability of ras to transform normal cells to cancer cells. (see e.g.
EP 0 461 869 A2, EP 0 496 162 A2, EP 0 523 873, and EP 0 520 823).
Thus it is believed that intracellular delivery of peptides having
the CAAX motif to transformed cells would be an effective
anti-neoplastic therapy.
Generally, however, small linear peptides do not make good
therapeutics because of their susceptability to proteolysis,
oxidation, and lack of transportability across cell membranes.
Accordingly, a need exists for a stable and potent non-peptidyl
farnesyl:protein transferase inhibitor that is permeable to cell
membranes.
Recently, several non-peptidyl ras farnesyl transferase inhibitors
were identified through microbial screening. Several antibiotics
(UCF1-A through UCF1-C) structurally related to manumycin inhibited
growth of Ki-ras-transformed fibrosarcoma [Hara, M., et al. Proc.
Natl, Sci. USA, 90:2281-2285 (1993)]. ##STR2## These inhibitors are
reported to have potential application in cancer therapy.
Burk, R., et al. WO 92/20336(Merck) also describe nonpeptidyl
farnesyltransferase inhibitors prepared by modification of natural
products having structures similar to the following compound:
##STR3## These compounds are reported to be useful in treating
cancer, especially colorectal carcinoma, exocrine pancreatic
carcinoma, and myloid leukemia.
Benzodiazepines and analogs thereof have been widely exploited as
therapeutics, but have not been reported to be inhibitors of
farnesylation of G-proteins such as p21.sup.ras. Benzodiazepines
are well known as central nervous system (CNS) drugs effecting the
neuro-inhibitory postsynaptic GABA receptor and chloride ionophore
channel (see eg. Watjen et al., J. Med. Chem. 32:2282-2291]1989]).
Benzodiazepine analogs have been employed as intermediates in the
synthesis of various anti-HIV-1 compounds [see e.g. Kukla, M. J. et
al., J. Med. Chem. 34:3187-3197 (1991)] and as antagonists of
gastrin and cholecystokinin (CCK) [see e.g. EP 0 284 256, assigned
to Merck, and Friedinger, Med. Res, Rev., 9 271 (1989)]. More
recently, benzodiazepine analogs have been reported to be
fibrinogen antagonists, inhibiting platelet aggregation [see e.g.
WO 93/00095 assigned to SmithKline Beecham.]
It was therefore an object of this invention to identify
nonpeptidyl compounds that more effectively antagonize
farnesylation of low molecular weight G-proteins such as
p21.sup.ras in disease states in animals, preferably mammals, and
especially humans. It was a further object of this invention to
identify compounds that inhibit isoprenylation of proteins in
microorganisms, such as yeast and fungi, that produce disease
states in plants or animals, preferably mammals, and especially
humans. These and other objects of this invention will be apparent
from consideration of the specification and claims as a whole.
SUMMARY OF THE INVENTION
The objects of this invention are accomplished by providing a
nonpeptidyl compound represented by structural formula (A):
##STR4## where
T.sup.1 is selected from CR.sup.1 R.sup.1', CR.sup.1, NR.sup.1, N,
O, and S(O).sub.u, where u is 0, 1, or 2;
T.sup.2 is selected from CR.sup.2 R.sup.2', CR.sup.2, NR.sup.2 and
N;
T.sup.3 is selected from CR.sup.3 R.sup.3', CR.sup.3, N, and
NR.sup.3 ;
R.sup.1, R.sup.1', R.sup.2, R.sup.2', R.sup.3 and R.sup.3' are
independently selected from hydrogen, C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl, halo(F, Cl, Br, I),
cyclohexyl, cyclohexenyl, phenyl, benzyl, and diphenylmethyl where
any phenyl moiety may be substituted with R or R', provided no halo
is bonded to any nitrogen;
R.sup.1 /R.sup.1', R.sup.2 /R.sup.2', and R.sup.3 /R.sup.3' each
pair taken together may independently form oxo (.dbd.O), provided
R.sup.1 /R.sup.1', and R.sup.2 /R.sup.2 are not simultaneously
.dbd.O;
R.sup.4 and R.sup.4' are independently selected from hydrogen,
hydroxy, C.sub.1 -C.sub.6 alkyl, halo(F, Cl, Br, I), halo(F, Cl,
Br, I)C.sub.1 -C.sub.6 alkyl, phenyl, and substituted phenyl where
the substituents are selected from halo(F, Cl, Br, I), halo(F, Cl,
Br, I)C.sub.1 -C.sub.6 alkyl, C.sub.1 -C.sub.6 alkoxy, and
nitro;
R.sup.5 and R.sup.6 are independently selected from hydrogen,
C.sub.1 -C.sub.6 alkyl, halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl,
phenyl, diphenylmethyl and substituted phenyl where the
substituents are selected from halo(F, Cl, Br, I) and nitro,
optionally, R.sup.5 and R.sup.6 together with the carbons to which
they are bonded may form a fused ring represented by ##STR5##
R.sup.5, R.sup.6, and R.sup.1 together with the carbons to which
they are bonded may form a fused ring system represented by
##STR6##
R and R' are one to three optional groups independently selected
from hydrogen, halo(F, Cl, Br, I), cyano, carboxamido,
carbamoyloxy, carboxyC.sub.1 -C.sub.12 alkyl, formyloxy, formyl,
azido, nitro, ureido, thioureido, hydroxy, mercapto, sulfonamido,
and an optionally substituted radical selected from C.sub.1
-C.sub.12 alkyl, C.sub.2 -C.sub.12 alkenyl, C.sub.3 -C.sub.12
alkynyl, C.sub.3 -C.sub.12 cycloalkyl, C.sub.6 -C.sub.14 aryl,
C.sub.6 -C.sub.10 aryl-C.sub.1 -C.sub.8 alkyl, C.sub.1 -C.sub.12
alkyloxy, C.sub.6 -C.sub.14 aryloxy, and C.sub.1 -C.sub.12
alkanoylamino, where the substituents are selected from halo(F, Cl,
Br, I), cyano, azido, nitro, hydroxy, mercapto, sulfonamido,
ureido, thioureido, carboxamido, carbamoyloxy, formyloxy, formyl,
C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, phenyl, and
phenoxy;
X is the side chain of any .alpha.-amino acid preferably an
L-.alpha.-amino acid and preferably selected from the group
--NR.sup.24 --C(.dbd.O)--R.sup.25, --NR.sup.24
--C(.dbd.O)--R.sup.8, --NR.sup.24 -- C(.dbd.O)NR.sup.7' R.sup.8,
--NR.sup.24 --C(.dbd.O)O--R.sup.8, --NR.sup.24
--C(.dbd.O)S--R.sup.8, --(CH.sub.2).sub.1-4 --NR.sup.24
--C(.dbd.O)--R.sup.25, --(CH.sub.2).sub.1-4 --C(.dbd.O)--R.sup.25,
--(CH.sub.2).sub.1-4 --C(.dbd.O)NH--R.sup.25, --(CH.sub.2).sub.0-4
--NR.sup.24 --CH(OH--R.sup.25, --CHR.sup.24 phenyl-R.sup.25,
--CHR.sup.24 phenoxy-R.sup.25, --CHR.sup.24 --O--R.sup.25,
--(CH.sub.2).sub.0-4 --NR.sup.24 --CH.sub.2 --R.sup.25,
--(CH.sub.2).sub.0-4 --NR.sup.24 --S(O).sub.u --R.sup.25 where u is
0, 1, or 2, --CHR.sup.24 --CH.sub.2 R.sup.25, --CHR.sup.24
--R.sup.25, --CR.sup.24 =CHR.sup.25 (E or Z), --(CH.sub.2).sub.0-4
--C.sub.6 -C.sub.10 aryl-R.sup.25, --(CH.sub.2).sub.0-4
-heterocycle-R.sup.25, --C.sub.1 -C.sub.2 haloalkyl-C.sub. 6
-C.sub.10 aryl-R.sup.25, and --C.sub.1 -C.sub.2
haloalkyl-heterocycle-R.sup.25, where any heterocycle is a 5- or
6-member saturated or unsaturated ring containing 1 to 3
heteroatoms selected from O, N, and S;
X together with the carbon to which it is bound and T.sup.2 may
form a heterocycle, where the heterocycle is a 5 or 6-member
saturated or unsaturated fused ring having from 1-3 hetero atoms
selected from O, N, and S, where any carbon atom of the heterocycle
is optionally substituted with oxo (.dbd.O) or R,
Y.sup.1, Y.sup.2, Y.sup.3, Y.sup.4 and Y.sup.5 are independently
selected from CH, CR, CR', and N;
Z is S or O;
W is selected from the group --C(.dbd.O)--NR.sup.7' R.sup.8,
--C(.dbd.O)--O--R.sup.8, --CR.sup.8' (OH)--CHR.sup.7 R.sup.8,
--CHR.sup.8' --CHR.sup.7 R.sup.8, --CR.sup.8' =CR.sup.7 R.sup.8 (E
or Z), --C(.dbd.O)--CHR.sup.7 R.sup.8, --CHR.sup.8' --NR.sup.7'
R.sup.8, --CHR.sup.8' --O--R.sup.8, --CHR.sup.8' --S(O).sub.u
--R.sup.8 where u is 0, 1, or 2; --CR.sup.8' =N--R.sup.8,
--CHR.sup.8' --R.sup.8, --C.sub.6 -C.sub.12 aryl-W', --C.sub.6
-C.sub.12 aryl-C.sub.1 -C.sub.3 alkyl-W', -heterocycle-W',
-heterocycle-C.sub.1 -C.sub.3 alkyl-W', -C.sub.1 -C.sub.2
alkyl-C.sub.6 -C.sub.10 aryl-W', and -C.sub.1 -C.sub.2
alkyl-heterocycle-W', where any heterocycle is a 5- or 6-member
saturated or unsaturated ring containing 1 to 3 heteroatoms
selected from O, N, and S;
W/R.sup.4' together with W/R.sup.3' and the carbon atoms to which
they are bound may form heterocycle-W' or heterocycle-C.sub.1
-C.sub.6 alkyl -W', where the heterocycle is a 5- or 6-member
saturated or unsaturated ring containing 1 to 3 heteroatoms
selected from O, N, and S and where the heterocycle is
unsubstituted or substituted with one or two substituents selected
from the group (i) --OH, (ii) --SH, (iii) --(C.sub.1 -C.sub.4
alkyl), (iv) --C.sub.1 -C.sub.4 alkoxyl, (v) CF.sub.3, (vi) halo(F,
Cl, Br, I), (vii) NO.sub.2, (viii) --COOH, (ix) --COO--(C.sub.1
-C.sub.4 alkyl), (x) --NH.sub.2, (xi) --NH(C.sub.1 -C.sub.4 alkyl),
and (xii) --N(C.sub.1 -C.sub.4 alkyl).sub.2 ;
W' is selected from one to three substituents selected from the
group hydrogen, --SR.sup.9, --SSR.sup.9, SC(.dbd.O)--R.sup.9,
--OR.sup.9, --C(.dbd.NH)--NH.sub.2, --N.dbd.CH--NH.sub.2,
--NH--CH.dbd.NH, R.sup.8, and V;
R.sup.7 is independently selected from the group hydrogen, C.sub.1
-C.sub.4 alkyl, halo(F, Cl, Br, I), and halo(F, Cl, Br, I)C.sub.1
-C.sub.4 alkyl;
R.sup.7 and X together may form ##STR7## where ##STR8## represents
a heterocycle bonded to the benzodiazepine moiety through a spiro
linkage, where the heterocycle is a 5- or 6-member saturated or
unsaturated nitrogen containing ring having from 0 to 2 additional
heteroatoms selected from O, N, and S, the ring optionally
containing a keto [--C(.dbd.O)--] group; and where X' is selected
from the group C(.dbd.O)--R.sup.25, CH(OH)--R.sup.25, CHR.sup.24
--R.sup.25, S(O).sub.u --R.sup.25 where u is 0, 1, or 2, CHR.sup.24
--R.sup.25, R.sup.25, C.sub.6 -C.sub.10 aryl-R.sup.25,
heterocycle-R.sup.25, C.sub.1 -C.sub.2 alkyl-C.sub.6 -C.sub.10
aryl-R.sup.25, and C.sub.1 -C.sub.2 alkyl-heterocycle-R.sup.25,
where any heterocycle is a 5- or 6-member saturated or unsaturated
ring containing 1 to 3 heteroatoms selected from O, N, and S;
R.sup.7' and R.sup.8' are selected from the group hydrogen, C.sub.1
-C.sub.4 alkyl, and halo(F, Cl, Br, I)C.sub.1 -C.sub.4 alkyl;
R.sup.7' and R.sup.8 together with the nitrogen to which they are
bonded may form a heterocyclic 5-, 6-, or 7-member ring containing
0, 1, or 2 additional heteroatoms selected from N, S, and O,
optionally substituted with one or two groups selected from
oxo(.dbd.O), --SR.sup.9, --SSR.sup.9, SC(.dbd.O)--R.sup.9,
--OR.sup.9, --C(.dbd.O)NHOH, --NHR.sup.9, --C(.dbd.O)NR.sup.27
R.sup.28, and --V;
R.sup.8' together with R.sup.3 and R.sup.8' together with R.sup.3'
may independently form a divalent radical selected from .dbd.CH--,
--CH.dbd., --CH.sub.2 --, --CH.sub.2 --CH.sub.2 --, .dbd.CH.sub.2
--CH.sub.2 --, --CH.dbd.CH--, and --CH.sub.2 --CH.dbd.;
R.sup.8 is selected from the group hydrogen, C.sub.1 -C.sub.8
alkyl, C.sub.1 -C.sub.4 alkyl-Z--C.sub.1 -C.sub.4 alkyl, where Z is
S or O, C.sub.2 -C.sub.4 alkyl-NR--C.sub.2 C.sub.4 alkyl, C.sub.2
-C.sub.8 alkenyl, C.sub.6 -C.sub.12 arylC.sub.1 -C.sub.3 alkyl,
indol-3-yl-C.sub.1 -C.sub.3 alkyl, and imidazol-4-yl-C.sub.1
-C.sub.3 alkyl, where any aryl moiety is optionally substituted
with --OR.sup.9 and V, and where any alkyl or alkenyl group is
optionally substituted with one to three groups selected from
--SR.sup.9, --SSR.sup.9, SC(.dbd.O)--R.sup.9, --OR.sup.9,
--C(.dbd.NH)--NH.sub.2, --N.dbd.CH--NH.sub.2, --NH--CH.dbd.NH,
--NH--C(.dbd.NH)--NH.sub.2, --C(.dbd.O)NHOH, --NHR.sup.9,
--C(.dbd.O)NR.sup.27 R.sup.28, and V;
V is selected from (a) --COR.sup.10, (b) --SO.sub.3 R.sup.13, (c)
--NHSO.sub.2 CF.sub.3, (d) --PO(OR.sup.13).sub.2, (e) --SO.sub.2
NHR.sup.10, (f) --CONHOR.sup.13, (g) --C(OH)R.sup.10
PO(OR.sup.13).sub.2, (h) --CN, (i) --SO.sub.2 NH-heteroaryl where
the heteroaryl is a 5- or 6-member aromatic ring containing 1 to 3
heteroatoms selected from O, N, and S and where the heteroaryl is
unsubstituted or substituted with one or two substituents selected
from the group (i) --OH, (ii) --SH, (iii) --(C.sub.1 -C.sub.4
alkyl), (iv) --C.sub.1 -C.sub.4 alkoxyl, (v) CF.sub.3, (vi) halo(F,
Cl, Br, I), (vii) NO.sub.2, (viii) --COOH, (ix) --COO--(C.sub.1
-C.sub.4 alkyl), (x) --NH.sub.2, (xi) --NH(C.sub.1 -C.sub.4 alkyl),
and (xii) --N(C.sub.1 -C.sub.4 alkyl).sub.2, (j) --CH.sub.2
SO.sub.2 -heterocycle, (k) --SO.sub.2 NHCOR.sup.10, (l) --CH.sub.2
SO.sub.2 NHCOR.sup.10, (m) --CONHSO.sub.2 R.sup.15, (n) --CH.sub.2
CONHSO.sub.2 R.sup.15, (o) --NHCONHSO.sub.2 R.sup.15, (p)
--NHSO.sub.2 NHCOR.sup.15, (q) --CONHNHSO.sub.2 CF.sub.3, (r)
CON(OH)R.sup.13, (s) --CONHCOCF.sub.3, (t) --CONHSO.sub.2 R.sup.10,
(u) --CONHSO.sub.2 R.sup.11, (v) --CONHSO.sub.2 R.sup.13,
##STR9##
R.sup.9 is selected from hydrogen, methyl, ethyl, isopropyl,
phenyl, and benzyl;
R.sup.10 is selected from the group consisting of (a) hydroxy, (b)
C.sub.1 -C.sub.8 -alkoxy, (c) C.sub.3 -C.sub.12 -alkenoxy, (d)
C.sub.6 -C.sub.12 -aryloxy, (e) C.sub.1 -C.sub.6 -alkyl-C.sub.6
-C.sub.12 -aryloxy, (f) di-C.sub.1 -C.sub.8 -alkylamino-C.sub.1
-C.sub.8 -alkoxy, (g) alkanoylamino-C.sub.1 -C.sub.8 -alkoxy
selected from the group (i) acetylaminoethoxy, (ii)
nicotinoylaminoethoxy, and (iii) succinamidoethoxy, (h) C.sub.1
-C.sub.8 -alkanoyloxy-C.sub.1 -C.sub.8 -alkoxy, (i) C.sub.6
-C.sub.12 -aryl-C.sub.1 -C.sub.8 -alkoxy where the aryl group is
unsubstituted or substituted with one to three of the groups (i)
nitro, (ii) halo (F, Cl, Br, I), (iii) C.sub.1 -C.sub.4 -alkoxy,
and (iv) amino, (j) hydroxy-C.sub.2 -C.sub.8 -alkoxy, (k)
dihydroxy-C.sub.3 -C.sub.8 -alkoxy, and (l) NR.sup.11 R.sup.12
;
R.sup.11 and R.sup.12 are independently selected from the group
consisting of (a) hydrogen, (b) C.sub.1 -C.sub.6 alkyl, (c) C.sub.2
-C.sub.6 alkanoyl, (d) C.sub.1 -C.sub.6 alkanoyl unsubstituted or
substituted with one to three of the groups (i) nitro, (ii) halo
(F, Cl, Br, I), (iii) C.sub.1 -C.sub.4 -alkoxy, and (iv) amino, and
(e) C.sub.6 -C.sub.12 -aryl-C.sub.1 -C.sub.8 -alkyl where the aryl
group is unsubstituted or substituted with one to three of the
groups (i) nitro, (ii) halo (F, Cl, Br, I), and (iii) C.sub.1
-C.sub.4 -alkoxy;
R.sup.13 is selected from the group consisting of (a) H, (b)
C.sub.1 -C.sub.6 alkyl, (c) halo(F, Cl, Br, I)-C.sub.1 -C.sub.6
alkyl, (d) phenyl, (e) benzyl, and (f) --CH.sub.2 --O--COCH.sub.3
;
R.sup.14 is selected from the group consisting of (a) H, (b) benzyl
and (c) --CH(R.sup.17)--O--C(O)R.sup.17 ;
R.sup.15 is selected from the group consisting of (a) C.sub.6
-C.sub.14 -aryl, (b) heteroaryl, (c) (C.sub.3 -C.sub.7)-cycloalkyl,
(d) (C.sub.1 -C.sub.4)-alkyl, unsubstituted or substituted with a
substituent selected from the group consisting of (i) aryl, (ii)
heteroaryl, (iii) --OH, (iv) --SH, (v) (C.sub.1 -C.sub.4)-alkyl,
(vi) (C.sub.1 -C.sub.4)-alkoxy, (vii) (C.sub.1 -C.sub.4)-alkylthio,
(viii) --CF.sub.3, (ix) halo (F, Cl, Br, I), (x) --NO.sub.2, (xi)
--CO.sub.2 H, (xii) CO.sub.2 --(C.sub.1 -C.sub.4)-alkyl, (xiii)
--NH.sub.2, (xiv) --N[(C.sub.1 -C.sub.4)-alkyl].sub.2, (xv)
--NH[(C.sub.1 -C.sub.4)-alkyl], (xvi) PO.sub.3 H and (xvii)
PO(OH)(C.sub.1 -C.sub.4)-alkoxy, and (e) (C.sub.1
-C.sub.4)-perfluoroalkyl;
R.sup.16 is selected from the group consisting of (a) --CN, (b)
--NO.sub.2, (c) --COOR.sup.13, (d) C.sub.1 -C.sub.6
-perfluoroalkyl, and (e) CF.sub.3 ;
R.sup.17 is independently selected from the group consisting of (a)
H, (b) (C.sub.1 -C.sub.6)-alkyl, (C.sub.2 -C.sub.6)-alkenyl,
(C.sub.2 -C.sub.6)-alkynyl or (C.sub.3 -C.sub.8)-cycloalkyl, each
of which is unsubstituted or substituted with (i) OH, (ii) (C.sub.1
-C.sub.4)-alkoxy, (iii) CO.sub.2 R.sup.15, (iv) OCOR.sup.15, (v)
CONHR.sup.15, (vi) CON(R.sup.15).sub.2, (vii)
N(R.sup.15)C(O)R.sup.15, (viii) NH.sub.2, (ix) (C.sub.1
-C.sub.4)-alkylamino, (x) di[(C.sub.1 -C.sub.4)-alkyl]amino, (xi)
aryl, and (xii) heteroaryl, (c) --C(O)-aryl, (d)-NO.sub.2, (e)
halo(Cl, Br, I, F), (f) --OH, (g) --OR.sup.18, (h) (C.sub.1
-C.sub.4)-perfluoroalkyl, (i) --SH, (j) --S(O).sub.1-2 (C.sub.1
-C.sub.4)-alkyl, (k) CO.sub.2 R.sup.15, (l) --SO.sub.3 H, (m)
--NR.sup.15 R.sup.18, (n) --NR.sup.15 C(O)R.sup.18, (o) --NR.sup.15
COOR.sup.14, (p) --SO.sub.2 NHR.sup.14, (q) --SO.sub.2 NR.sup.15
R.sup.15, (r) --NHSO.sub. 2 R.sup.14, (s) --C(O)NHSO.sub.2
R.sup.14, (t) aryl, (u) heteroaryl, (v) morpholin-4-yl, (w)
CONH.sub.2, and (y) 1H-tetrazol-5-yl;
R.sup.18 is selected from the group consisting of (a) H and (b)
(C.sub.1 -C.sub.4)-alkyl unsubstituted or substituted with (i)
NH.sub.2, (ii) NH[(C.sub.1 -C.sub.4)-alkyl], (iii) N[(C.sub.1
-C.sub.4)-alkyl].sub.2, (iv) CO.sub.2 H, (v) CO.sub.2 (C.sub.1
-C.sub.4)-alkyl, (vi) OH, (vii) SO.sub.3 H, and (viii) SO.sub.2
NH.sub.2 ;
R.sup.19 is selected from the group consisting of (a) H, (b)
(C.sub.1 -C.sub.6)-alkyl, (c) (C.sub.2 -C.sub.6)-alkenyl, (d)
(C.sub.1 -C.sub.6)-alkoxy, (e) (C.sub.2 -C.sub.6)-alkoxyalkyl, (f)
--CH.sub.2 --O--COCH.sub.3, or (g) benzyl, where the phenyl moiety
is unsubstituted or substituted with a group selected from
--NO.sub.2, --NH.sub.2, --OH, or --OCH.sub.3 ;
R.sup.20, R.sup.21, and R.sup.22 are each independently selected
from H, Cl, CN, NO.sub.2, CF.sub.3, C.sub.2 F.sub.5, C.sub.3
F.sub.7, CHF.sub.2, CH.sub.2 F, CO.sub.2 CH.sub.3, CO.sub.2 C.sub.2
H.sub.5, SO.sub.2 CH.sub.3, SO.sub.2 CH.sub.3, SO.sub.2 CF.sub.3
and SO.sub.2 C.sub.6 F.sub.5, where Q is selected from O, S,
NR.sup.23 and CH.sub.2 ;
R.sup.23 is selected from hydrogen, CH.sub.3, and CH.sub.2 C.sub.6
H.sub.5 ;
R.sup.24 is selected from hydrogen, C.sub.1 -C.sub.6 alkyl, benzyl,
halo(F, Cl, Br, I)benzyl, and halo(F, Cl, Br, I)C.sub.1 -C.sub.6
alkyl;
R.sup.24 and R.sup.7 together may form an ethylene, ethenylene,
propylene, propenylene, butylene, or butenylene bridge;
R.sup.25 is selected from R.sup.25', ##STR10##
R.sup.25' is selected from --SR.sup.26, --SSR.sup.26, --OR.sup.26,
--NOR.sup.26, C.sub.1 -C.sub.6 alkyl, C.sub.2 -C.sub.6 alkenyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl where any alkyl or alkenyl
moiety is optionally substituted with one to three groups selected
from --SR.sup.26, --SSR.sup.26, --OR.sup.26, --NOR.sup.26 and
--NR.sup.27 R.sup.28 ;
R.sup.26 is selected from hydrogen, C.sub.1 -C.sub.6 alkyl, halo(F,
Cl, Br, I)C.sub.1 -C.sub.6 alkyl, and C.sub.1 -C.sub.6
alkanoyl;
R.sup.27 and R.sup.28 are independently selected from hydrogen,
C.sub.1 -C.sub.6 alkyl, phenyl, napthyl, benzyl, --CH.sub.2
-napthyl, C.sub.1 -C.sub.6 alkanoyl, C.sub.2 -C.sub.6 alkanoyl
optionally substituted with --COOH and --NH.sub.2, C.sub.1 -C.sub.6
cycloalkanoyl. C.sub.6 -C.sub.10 aroyl, C.sub.6 -C.sub.10
arylC.sub.1 -C.sub.6 alkanoyl, C.sub.1 -C.sub.6 alkylsulfonyl,
C.sub.6 -C.sub.10 arylsulfonyl, C.sub.6 -C.sub.10 arylC.sub.1
-C.sub.6 alkylcarbamoyl, cinnamoyl, heterocyclecarbonyl, C.sub.1
-C.sub.6 alkoxycarbonyl, C.sub.6 -C.sub.10 aryloxycarbonyl, C.sub.6
-C.sub.10 arylC.sub.1 -C.sub.6 alkoxycarbonyl, and
pyroglutamyl;
R.sup.27 and R.sup.24 together may form a diradical selected from
--CH.sub.2 --, --C(.dbd.O)--, --CH.sub.2 --CH.sub.2 --, and
--CH.sub.2 --C(.dbd.O)--;
R.sup.27 and R.sup.28 together with the nitrogen atom to which they
are bonded may form ##STR11## or a cyclic imide represented by
##STR12##
G is selected from --CH.sub.2 --, O, S(O).sub.u where u is 0, 1, or
2, and NR.sup.28 ;
J-M is selected from C.sub.2 -C.sub.4 alkylene and C.sub.2 -C.sub.4
alkenylene;
R.sup.29 is selected from hydrogen, C.sub.1 -C.sub.6 alkyl, halo(F,
Cl, Br, I)C.sub.1 -C.sub.6 alkyl, phenyl, benzyl, and pyridyl where
any phenyl moiety may be substituted with halo(F and Cl),
--CF.sub.3, --NO.sub.2, --NH.sub.2, --OH, and --OCH.sub.3 ; and
pharmaceutically acceptable salts thereof.
Preferably the compounds of this invention are selected from those
represented by structural formulae I-IX: ##STR13## where R, R',
R.sup.1, R.sup.4, R.sup.4', R.sup.7, R.sup.7', R.sup.8, R.sup.8',
R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.15,
R.sup.16, R.sup.19, R.sup.24, R.sup.25, R.sup.25', R.sup.26,
R.sup.27, R.sup.28, R.sup.29, W, W', V, X, X', G, and J-M are
defined above;
Optionally, R.sup.1 and R.sup.2 taken together may form a covalent
bond or fused benzene substituted with R and R'; ##STR14##
represents a heterocycle bonded to the benzodiazepine moiety
through a spiro linkage, where the heterocycle is a 5- or 6-member
saturated or unsaturated nitrogen containing ring having from 0 to
2 additional heteroatoms selected from O, N, and S, the ring
optionally containing a keto [--C(.dbd.O)--] group; ##STR15##
represents a heterocycle fused to the benzodiazepine moiety, where
the heterocycle is a 5- or 6-member saturated or unsaturated
di-nitrogen containing ring having from 0 to 1 additional
heteroatom selected from O, N, and S, the ring optionally
containing a keto [--C(.dbd.O)--] group; ##STR16## represents a
heterocycle bonded to the benzodiazepine moiety through a ring
nitrogen, where the heterocycle is a 5- or 6-member saturated or
unsaturated nitrogen containing ring having from 0 to 2 additional
heteroatoms selected from O, N, and S, the ring optionally
containing a keto [--C(.dbd.O)--] group; ##STR17## represents
C.sub.6 -C.sub.10 aryl or a heteroaryl where the heteroaryl is a 5-
or 6-member aromatic ring containing 1 to 3 heteroatoms selected
from O, N, and S, the C.sub.6 -C.sub.10 aryl or heteroaryl is
optionally substituted with V and R.sup.8 ;
A.sup.1 and A.sup.2 are independently selected from CRR', CR',
CRR.sup.8, CR.sup.8, N, O, and S provided one of A.sup.1 and
A.sup.2 is CRR.sup.8 or CR.sup.8 ; and ##STR18## represents a
single or double bond.
Most preferred compounds of the instant invention are represented
by formulae (Ia)-(Ie): ##STR19##
R and R' are independently selected from the group hydrogen,
halo(F, Cl, Br, I), halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl, and
C.sub.1 -C.sub.6 alkoxy;
R.sup.4 and R.sup.4' are independently selected from hydrogen,
halo(F, Cl, Br, I), C.sub.1 -C.sub.6 alkyl, and halo(F, Cl, Br,
I)C.sub.1 -C.sub.6 alkyl,
R.sup.7 is hydrogen;
R.sup.7' is selected from the group hydrogen, C.sub.1 -C.sub.4
alkyl, and halo(F, Cl, Br, I)C.sub.1 -C.sub.4 alkyl;
R.sup.8 is selected from the group unsubstituted and substituted
C.sub.1 -C.sub.8 alkyl, phenyl-C.sub.1 -C.sub.3 alkyl,
indol-3-yl-C.sub.1 -C.sub.3 alkyl, and imidazol-4-yl-C.sub.1
-C.sub.3 alkyl, where any phenyl moiety is optionally substituted
with --OR.sup.9 and where any alkyl group is optionally substituted
with one or two groups selected from --SR.sup.9, --SSR.sup.9,
--SC(.dbd.O)--R.sup.9, --OR.sup.9, --C(.dbd.O)NHOH, --NHR.sup.9,
--C(.dbd.O)NR.sup.27 R.sup.28, and --V;
R.sup.9 is selected from hydrogen, methyl, ethyl, isopropyl,
phenyl, and benzyl;
V is selected from the group --COR.sup.10, and ##STR20##
R.sup.10 is selected from the group hydroxy, and C.sub.1 -C.sub.8
-alkoxy;
R.sup.19 is selected from hydrogen, C.sub.1 -C.sub.6 -alkyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 -alkyl;
X is selected from the group --NR.sup.24 --C(.dbd.O)--R.sup.25,
--NR.sup.24 --C(.dbd.O)--R.sup.8, --NR.sup.24 --C(.dbd.O)NR.sup.7'
R.sup.8, --NR.sup.24 --CH(OH)--R.sup.25, --NR.sup.24 --CH.sub.2
--R.sup.25, --NR.sup.24 --S(O).sub.u --R.sup.25 where u is 0, 1, or
2, --CHR.sup.24 --CH.sub.2 R.sup.25, --CHR.sup.24 --R.sup.25,
--CR.sup.24 .dbd.CHR.sup.25 (E or Z), --C.sub.6 -C.sub.10
aryl-R.sup.25, -heterocycle-R.sup.25, --C.sub.1 -C.sub.2
alkyl-C.sub.6 -C.sub.10 aryl-R.sup.25, and --C.sub.1 -C.sub.2
alkyl-heterocycle-R.sup.25, where any heterocycle is a 5- or
6-member saturated or unsaturated ring containing 1 to 3
heteroatoms selected from O, N, and S;
R.sup.24 is selected from C.sub.1 -C.sub.6 alkyl, benzyl, and
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl;
R.sup.25 is selected from R.sup.25', ##STR21##
R.sup.25' is selected from --SR.sup.26, --SSR.sup.26, --OR.sup.26,
--(C.dbd.O)NOR.sup.26, C.sub.1 -C.sub.6 alkyl, C.sub.6 -C.sub.12
arylC.sub.1 -C.sub.6 alkyl, C.sub.2 -C.sub.6 alkenyl, C.sub.1
-C.sub.6 alkylamine, C.sub.2 -C.sub.6 alkenylamine, and halo(F, Cl,
Br, I)C.sub.1 -C.sub.6 alkyl where any alkyl or alkenyl moiety is
optionally substituted with --SR.sup.26, --SSR.sup.26, --OR.sup.26,
--(C.dbd.O)NOR.sup.26 and --NR.sup.27 R.sup.28, and where any amine
moiety is optionally substituted with R.sup.27 or R.sup.28 ;
R.sup.25" is selected from hydrogen, C.sub.1 -C.sub.6 alkyl,
halo(F, Cl, Br, I)C.sub.1 -C.sub.6 alkyl, C.sub.6 -C.sub.12 aryl,
C.sub.6 -C.sub.12 arylC.sub.1 -C.sub.6 alkyl, where any alkyl or
aryl moiety may optionally be substituted with a group selected
from --SR.sup.26, --SSR.sup.26, --OR.sup.26, COR.sup.10, and
NOR.sup.26 ;
R.sup.26 is selected from hydrogen, C.sub.1 -C.sub.6 alkyl, halo(F,
Cl, Br, I)C.sub.1 -C.sub.6 alkyl, and C.sub.1 -C.sub.6
alkanoyl;
R.sup.27 and R.sup.28 are independently selected from hydrogen,
C.sub.1 -C.sub.6 alkyl, phenyl, napthyl, benzyl, --CH.sub.2
-napthyl, C.sub.1 -C.sub.6 alkanoyl, C.sub.1 -C.sub.6
cycloalkanoyl. C.sub.6 -C.sub.10 aroyl, C.sub.6 -C.sub.10
arylC.sub.1 -C.sub.6 alkanoyl, C.sub.1 -C.sub.6 alkylsulfonyl,
C.sub.6 -C.sub.10 arylsulfonyl, C.sub.6 -C.sub.10 arylC.sub.1
-C.sub.6 alkylcarbamoyl, cinnamoyl, heterocyclecarbonyl, C.sub.1
-C.sub.6 alkoxycarbonyl, C.sub.6 -C.sub.10 aryloxycarbonyl, C.sub.6
-C.sub.10 arylC.sub.1 -C.sub.6 alkoxycarbonyl, and
pyroglutamyl;
R.sup.27 and R.sup.28 together with the nitrogen atom to which they
are bonded may form a cyclic amine represented by ##STR22##
G is selected from --CH.sub.2 --, O, S(O).sub.u where u is 0, 1, or
2, and NR.sup.28, and pharmaceutically acceptable salts
thereof.
The very most preferred compounds of this invention include;
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl] acetyl]-(D or L)-methionine
cyclohexyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl] acetyl]-(D or L)-methionine isopentyl
ester,
N-[[3-(2-
Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,
4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine morpholino-N-ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-
1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1
-yl]acetyl]-(D or L)-methionine methyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
cholesteryl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5oxo-5
-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
isopentyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine cholesteryl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine
cyclohexyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine
isopentyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine
cholesteryl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-phenylalanine
isobutyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine isopentyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
cholesteryl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine isopentyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
cholesteryl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine isopentyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine cholesteryl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine isopentyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine cholesteryl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine isobutyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan cyclohexyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan isopentyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan ethyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan methyl
ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
cholesteryl ester,
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan isobutyl
ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methioine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-methionine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl] acetyl]-(D or L)-isoleucine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-isoleucine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-norleucine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine ethyl
ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tyrosine
isobutyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
cyclohexyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
isopentyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
ethyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
methyl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
cholesteryl ester,
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-o
xo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-tryptophan
isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-methionine isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-14-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-leucine
isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-phenylalanine isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-isoleucine isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-norleucine isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl] acetyl]-(D or L)-valine
ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-
(D or L)-valine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or L)-valine
isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine ethyl ester,
2N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihyd
ro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tyrosine isobutyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan cyclohexyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan isopentyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan morpholino-N-ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan ethyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan methyl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan cholesteryl ester,
N-[[3-(2-Amino-3-tert-butylthiomercapto-1-oxopropyl)methylamino]-2,3-dihydr
o-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-(D or
L)-tryptophan isobutyl ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine cyclohexyl
ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine isopenyl ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine morpholinoethyl
ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine ethyl ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine methyl ester,
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine cholesteryl ester,
and
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)methylamino]-2,3,4,5-tetrahydro-
2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methionine isobutyl ester.
The invention provides a pharmaceutical composition comprising a
pharmaceutically acceptable excipient and the compound of formulae
A, I-IX, or IXa-IXd and a method of inhibiting farnesyl:protein
transferse comprising administering to a subject in need of such
treatment a therapeutically effective amount of the composition.
The invention also provides a method of inhibiting farnesylation of
the oncogene protein ras in a subject and a method of amelioration
of a neoplastic or proliferative condition in a subject having such
a condition comprising administering to the subject in need of such
treatment a therapeutically effective amount of the pharmaceutical
composition.
The invention further provides a method of inhibiting fungal growth
or reproduction in a living organism (or an area where growth or
reproduction is to be controlled) in need of such treatment
comprising administering an antifungally effective amount of the
compound of formulae A, I-IX, or IXa-IXd.
BRIEF DESCRIPTION OF THE FIGURES
FIGS. 1A-1C Differential inhibition of CAAX farnesyltransferase
(A), CAAX GG transferase (B) and Rab GG transferase (C) by compound
27 B (denoted as BZA-2B in the figure, open circles) and the
tetrapeptide Cys-Val-Phe-Met (CVFM) (solid circles).
FIGS. 2A and 2B Differential inhibition of CAAX farnesyltransferase
(A) and CAAX GG transferase (B) by compound 27B (denoted as BZA-2B
in the figure, closed circles) and compound 31B (denoted as BZA-4B
in the figure, open triangles).
FIG. 3 Inhibition of [3H]mevalonate incorporation into prenylated
proteins in monolayers of hamster Met18b-2 cells by compound 31B
(denoted as BZA-4B in the figure). Effects of control peptides
SVIM, CVIM and CVFM as well as compound 27A (BZA-2A in the figure)
and 31A (BZA-4A in the figure) are also shown.
FIG. 4 Dose dependence of inhibition of [3H]mevalonate
incorporation into prenylated proteins in monolayers of hamster
met18b-2 cells by compound 27B (denoted as BZA-2B in the figure)
and compound 37B (denoted as BZA-5B in the figure) and compound 33B
(denoted as BZA-6B in the figure).
FIG. 5 Inhibition of [3H]mevalonate incorporation into p21H-ras
proteins in Met18b-2 cells by compound 37B (denoted as BZA-5B in
the figure). Triton soluble fractions (lanes 1-5) and the same
fractions following immunoprecipitation with an anti-ras monoclonal
antibody (lanes 6-10) are also shown.
FIGS. 6A-6F Morphology of H-ras(Val12)-transformed rat-1
fibroblasts (A,B), src-transformed rat-1 firbroblasts (C,D), and
untrasformed rat-1 fibroblasts (E,F) incubated in the presence of
either compound 27A,
N-[[3-(2(S)-amino-3-mercapto-1-oxopropyl)methylamine]-2,3-dihydro-2-oxo-5-
phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (isomer A)
denoted BZA-2A in the figure (A,C,E) or compound 27B,
N-[[3-(2(S)-amino-3-mercapto-1-oxopropyl)methylamine]-2,3-dihydro-2-oxo-5-
phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (isomer B)
(B,D,F).
FIGS. 7A-7F Effects of farnesyltransferase inhibition on
ras-transformed rat-1 fibroblasts (A, B), src-transformed rat-1
fibroblasts (C, D), and untransformed parental rat-1 fibroblasts
(E, F). Growth rate of cell lines in the absence (A, C, E) and
presence (B, D, F) of the inhibitor BZA-5B is shown.
DETAILED DESCRIPTION OF THE INVENTION
A. Definitions
Terms used in the claims and specification are defined as set forth
below unless otherwise specified.
The term "alkyl" means a branched or unbranched, saturated
aliphatic hydrocarbon radical, having the number of carbon atoms
specified, or if no number is specified, having up to 12 carbon
atoms. Examples of alkyl radicals include methyl, ethyl, n-propyl,
isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl,
2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl,
2,2-dimethylbutyl, n-heptyl, 2-methylhexyl, and the like. The terms
"lower alkyl" and "C.sub.1 -C.sub.6 alkyl" are synonymous and used
interchangeably. A preferred "C.sub.1 -C.sub.6 alkyl" group is
methyl.
The term "substituted C.sub.n -C.sub.m alkyl" where m and n are
integers identifying the range of carbon atoms contained in the
alkyl group, denotes the above alkyl groups that are substituted by
one, two or three; halogen(F, Cl. Br, I), hydroxy, protected
hydroxy, amino, protected amino, C.sub.1 -C.sub.6 acyloxy, nitro,
carboxy, protected carboxy, carbamoyl, carbamoyloxy, cyano,
methylsulfonylamino or C.sub.1 -C.sub.6 alkoxy groups. The
substituted alkyl groups may be substituted once, twice or three
times with the same or with different substituents.
Examples of the above substituted alkyl groups include but are not
limited to; cyanomethyl, nitromethyl, hydroxymethyl,
trityloxymethyl, propionyloxymethyl, aminomethyl, carboxymethyl,
alkyloxycarbonylmethyl, allyloxycarbonylaminomethyl,
carbamoyloxymethyl, methoxymethyl, ethoxymethyl, t-butoxymethyl,
acetoxymethyl, chloromethyl, bromomethyl, iodomethyl,
trifluromethyl, 6-hydroxyhexyl, 2,4-dichloro(n-butyl),
2-amino(isopropyl), 2-carbamoyloxyethyl and the like. A preferred
group of examples within the above "C.sub.1 -C.sub.12 substituted
alkyl" group includes the substituted methyl group, e.g. a methyl
group substituted by the same substituents as the "substituted
C.sub.n -C.sub.m alkyl" group. Examples of the substituted methyl
group include groups such as hydroxymethyl, protected hydroxymethyl
(e.g. tetrahydropyranyloxymethyl), acetoxymethyl,
carbamoyloxymethyl, trifluromethyl, chloromethyl, bromomethyl and
iodomethyl.
The terms "C.sub.1 -C.sub.6 alkyloxy" or "C.sub.1 -C.sub.6 alkoxy"
are used interchangeably herein and denote groups such as methoxy,
ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy and like
groups.
The terms "C.sub.1 -C.sub.12 acyloxy" or "C.sub.1 -C.sub.12
alkanoyloxy" are used interchangeably and denote herein groups such
as formyloxy, acetoxy, propionyloxy, butyryloxy, pentanoyloxy,
hexanoyloxy, heptanoyloxy, and the like.
The terms "C.sub.1 -C.sub.12 alkylcarbonyl", "C.sub.1 -C.sub.12
alkanoyl" and "C.sub.1 -C.sub.12 acyl" are used interchangeably
herein encompass groups such as formyl, acetyl, propionyl, butyryl,
pentanoyl, hexanoyl, heptanoyl, benzoyl and the like.
The term "cycloalkyl" as used herein refers to a mono-, bi-, or
tricyclic aliphatic ring having 3 to 14 carbon atoms and preferably
3 to 7 carbon atoms.
The term "alkenyl" means a branched or unbranched hydrocarbon
radical having the number of carbon atoms designated containing one
or more carbon-carbon double bonds, each double bond being
independently cis, trans, or a nongeometric isomer.
The terms "C.sub.1 -C.sub.12 alkylthio" and "C.sub.1 -C.sub.12
substituted alkylthio" denote C.sub.1 -C.sub.12 alkyl and C.sub.1
-C.sub.12 substituted alkyl groups, respectively, attached to a
sulfur which is in turn the point of attachment for the alkylthio
or substituted alkylthio group to the group or substituent
designated.
The term "aryl" when used alone means a homocyclic hydrocarbon
aromatic radical, whether or not fused, having the number of carbon
atoms designated. Preferred aryl groups include phenyl, napthyl,
biphenyl, phenanthrenyl, naphthacenyl, and the like (see e.g.
Lang's Handbook of Chemistry (Dean, J. A., ed) 13.sup.th ed. Table
7-2 [1985]).
The term "substituted phenyl" or "substituted aryl" denotes a
phenyl group or aryl group substituted with one, two or three
substituents chosen from halogen(F, Cl, Br, I), hydroxy, protected
hydroxy, cyano, nitro, C.sub.1 -C.sub.6 alkyl, C.sub.1 -C.sub.6
alkoxy, carboxy, protected carboxy, carboxymethyl, protected
carboxymethyl, hydroxymethyl, protected hydroxymethyl, aminomethyl,
protected aminomethyl, trifluoromethyl N-(methylsulfonylamino) or
other groups specified.
Examples of the term "substituted phenyl" includes but is not
limited to a mono- or di(halo)phenyl group such as 4-chlorophenyl,
2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl,
3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl,
3-chloro-4-fluorophenyl, 2-fluorophenyl and the like; a mono- or
di(hydroxy)phenyl group such as 4-hydroxyphenyl, 3-hydroxyphenyl,
2,4-dihydroxyphenyl, the protected-hydroxy derivatives thereof and
the like; a nitrophenyl group such as 3- or 4-nitrophenyl; a
cyanophenyl group, for example, 4-cyanophenyl; a mono- or di(lower
alkyl)phenyl group such as 4-methylphenyl, 2,4-dimethylphenyl,
2-methylphenyl, 4-(iso-propyl)phenyl, 4-ethylphenyl,
3-(n-propyl)phenyl and the like; a mono or di(alkoxy)phenyl group,
for example, 2,6-dimethoxyphenyl, 4-methoxyphenyl, 3-ethoxyphenyl,
4-(isopropoxy)phenyl, 4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl
and the like; 3- or 4- trifluoromethylphenyl; a mono- or
dicarboxyphenyl or (protected carboxy)phenyl group such
4-carboxyphenyl; a mono- or di(hydroxymethyl)phenyl or (protected
hydroxymethyl)phenyl such as 3-(protected hydroxymethyl)phenyl or
3,4-di(hydroxymethyl)phenyl; a mono- or di(aminomethyl)phenyl or
(protected aminomethyl)phenyl such as 2-(aminomethyl)phenyl or
2,4-(protected aminomethyl)phenyl; or a mono- or
di(N-(methylsulfonylamino))phenyl such as
3-(N-methylsulfonylamino))phenyl. Also, the term "substituted
phenyl" represents disubstituted phenyl groups wherein the
substituents are different, for example, 3-methyl-4-hydroxyphenyl,
3-chloro-4-hydroxyphenyl, 2-methoxy- 4-bromophenyl,
4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,
2-hydroxy-4-chlorophenyl and the like. Preferred substituted phenyl
groups include the 2- and 3-trifluoromethylphenyl, the
4-hydroxyphenyl, the 2-aminomethylphenyl and the
3-(N-(methylsulfonylamino))phenyl groups.
The term "arylalkyl" means one, two, or three aryl groups having
the number of carbon atoms designated, appended to an alkyl radical
having the number of carbon atoms designated including but not
limited to; benzyl, napthylmethyl, phenethyl, benzyhydryl
(diphenylmethyl), trityl, and the like. A preferred arylalkyl group
is the benzyl group.
The term "substituted C.sub.6 -C.sub.12 aryl-C.sub.1 -C.sub.6
alkyl" denotes a C.sub.1 -C.sub.6 alkyl group substituted at any
carbon with a C.sub.6 -C.sub.12 aryl group bonded to the alkyl
group through any aryl ring position and substituted on the C.sub.1
-C.sub.6 alkyl portion with one, two or three groups chosen from
halogen (F, Cl, Br, I), hydroxy, protected hydroxy, amino,
protected amino, C.sub.1 -C.sub.6 acyloxy, nitro, carboxy,
protected carboxy, carbamoyl, carbamoyloxy, cyano, C.sub.1 -C.sub.6
alkylthio, N-(methylsulfonylamino) or C.sub.1 -C.sub.6 alkoxy.
Optionally the aryl group may be substituted with one, two, or
three groups chosen from halogen, hydroxy, protected hydroxy,
nitro, C.sub.1 -C.sub.6 alkyl, C.sub.1 -C.sub.4 alkoxy, carboxy,
protected carboxy, carboxymethyl, protected carboxymethyl,
hydroxymethyl, protected hydroxymethyl, aminomethyl, protected
aminomethyl, or an N-(methylsulfonylamino) group. As before, when
either the C.sub.1 -C.sub.6 alkyl portion or the aryl portion or
both are disubstituted, the substituents can be the same or
different.
Examples of the term "substituted C.sub.6 -C.sub.10 aryl-C.sub.1
-C.sub.6 alkyl" include groups such as 2-phenyl-1-chloroethyl,
2-(4-methoxyphenyl)ethyl, 2,6-dihydroxy-4-phenyl(n-hexyl),
5-cyano-3-methoxy-2-phenyl(n-pentyl),
3-(2,6-dimethylphenyl)n-propyl, 4-chloro-3-aminobenzyl,
6-(4-methoxyphenyl)-3-carboxy(n-hexyl), 5-(4-aminomethyl
phenyl)-3-(aminomethyl)(n-pentyl), and the like.
The term "carboxy-protecting group" as used herein refers to one of
the ester derivatives of the carboxylic acid group commonly
employed to block or protect the carboxylic acid group while
reactions are carried out on other functional groups on the
compound. Examples of such carboxylic acid protecting groups
include 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl,
2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl,
pentamethylbenzyl, 3,4-methylenedioxybenzyl, benzhydryl,
4,4'-dimethoxybenzhydryl, 2,2',4,4'-tetramethoxybenzhydryl,
t-butyl, t-amyl, trityl,4-methoxytrityl, 4,4'-dimethoxytrityl,
4,4',4"-trimethoxytrityl, 2-phenylprop-2-yl, trimethylsilyl,
t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,
.beta.-(trimethylsilyl)ethyl, .beta.-(di(n-butyl)methylsilyl)ethyl,
p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl,
cinnamyl, 1-(trimethylsilylmethyl)prop-1-en-3-yl, and like
moieties. The species of carboxy-protecting group employed is not
critical so long as the derivatized carboxylic acid is stable to
the condition of subsequent reaction(s) on other postions of the
benzodiazepinedione molecule and can be removed at the appropriate
point without disrupting the remainder of the molecule. In
particular, it is important not to subject the carboxy-protected
benzodiazepinedione molecule to strong nucleophilic bases or
reductive conditions employing highly activated metal catalysts
such as Raney nickel. (Such harsh removal conditions are also to be
avoided when removing amino-protecting groups and
hydroxy-protecting groups, discussed below.) Preferred carboxylic
acid protecting groups are the allyl and p-nitrobenzyl groups.
Similar carboxy-protecting groups used in the cephalosporin,
penicillin and peptide arts can also be used to protect a carboxy
group substituents of the benzodiazepinedione. Further examples of
these groups are found in E. Haslam, "Protective Groups in Organic
Chemistry", J. G. W. McOmie, Ed., Plenum Press, New York, N.Y.,
1973, Chapter 5, and T. W. Greene, "Protective Groups in Organic
Synthesis", John Wiley and Sons, New York, NY, 1981, Chapter 5. The
term "protected carboxy" refers to a carboxy group substituted with
one of the above carboxy-protecting groups.
As used herein the term "amide-protecting group" refers to any
group typically used in the peptide art for protecting the peptide
nitrogens from undesirable side reactions. Such groups include
p-methoxyphenyl, 3,4-dimethoxybenzyl, benzyl, O-nitrobenzyl,
di-(p-methoxyphenyl)methyl, triphenylmethyl,
(p-methoxyphenyl)diphenylmethyl, diphenyl-4-pyridylmethyl,
m-2-(picolyl)-N'-oxide, 5-dibenzosuberyl, trimethylsilyl, t-butyl
dimethylsilyl, and the like. Further descriptions of these
protecting groups can be found in "Protective Groups in Organic
Synthesis", by Theodora W. Greene, 1981, John Wiley and Sons, New
York.
Unless otherwise specified, the terms "heterocycle", "heterocyclic
group", "heterocyclic" or "heterocyclyl" are used interchangeably
herein and refer to any mono-, bi-, or tricyclic saturated,
unsaturated, or aromatic ring having the number of ring atoms
designated where at least one ring is a 5-, 6- or 7-membered
hydrocarbon ring containing from one to four heteroatoms selected
from nitrogen, oxygen, and sulfur, preferably at least one
heteroatom is nitrogen (Lang's Handbook of Chemistry, supra).
Preferably, the heterocycle is a 5- or 6-member saturated,
unsaturated, or aromatic hydrocarbon ring containing 1, 2, or 3
heteroatoms selected from O, N, and S. Typically, the 5-membered
ring has 0 to 2 double bonds and the 6- or 7-membered ring has 0 to
3 double bonds and the nitrogen or sulfur heteroatoms may
optionally be oxidized, and any nitrogen heteroatom may optionally
be quarternized. Included in the definition are any bicyclic groups
where any of the above heterocyclic rings are fused to a benzene
ring. Heterocyclics in which nitrogen is the heteroatom are
preferred.
The following ring systems are examples of the heterocyclic
(whether substituted or unsubstituted) radicals denoted by the term
"heterocylic": thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl,
thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl,
thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl,
pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, thiazinyl, oxazinyl,
triazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl,
oxathiazinyl, tetrazinyl, thiatriazinyl, oxatriazinyl,
dithiadiazinyl, imidazolinyl, dihydropyrimidyl,
tetrahydropyrimidyl, tetrazolo[1,5-b]pyridazinyl and purinyl, as
well as benzo-fused derivatives, for example benzoxazolyl,
benzthiazolyl, benzimidazolyl and indolyl.
Heterocyclic 5-membered ring systems containing a sulfur or oxygen
atom and one to three nitrogen atoms are also suitable for use in
the instant invention. Examples of such preferred groups include
thiazolyl, in particular thiazol-2-yl and thiazol-2-yl N-oxide,
thiadiazolyl, in particular 1,3,4-thiadiazol-5-yl and
1,2,4-thiadiazol-5-yl, oxazolyl, preferably oxazol-2-yl, and
oxadiazolyl, such as 1,3,4-oxadiazol-5-yl, and
1,2,4-oxadiazol-5-yl. A group of further preferred examples of
5-membered ring systems with 2 to 4 nitrogen atoms include
imidazolyl, preferably imidazol-2-yl; triazolyl, preferably
1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and
tetrazolyl, preferably 1H-tetrazol-5-yl. A preferred group of
examples of benzo-fused derivatives are benzoxazol-2-yl,
benzthiazol-2-yl and benzimidazol-2-yl.
Further suitable specific examples of the above heterocylic ring
systems are 6-membered ring systems containing one to three
nitrogen atoms. Such examples include pyridyl, such as pyrid-2-yl,
pyrid-3-yl, and pyrid-4-yl; pyrimidyl, preferably pyrimid-2-yl and
pyrimid-4-yl; triazinyl, preferably 1,3,4-triazin-2-yl and
1,3,5-triazin-4-yl; pyridazinyl, in particular pyridazin-3-yl, and
pyrazinyl. The pyridine N-oxides and pyridazine N-oxides and the
pyridyl, pyrimid-2-yl, pyrimid-4-yl, pyridazinyl and the
1,3,4-triazin-2-yl radicals, are a preferred group. Optionally
preferred 6-membered ring heterocycles are; piperazinyl,
piperazin-2-yl, piperidyl, piperid-2-yl, piperid-3-yl,
piperid-4-yl, morpholino, morpholin-2-yl, and morpholin-3-yl.
The substituents for the optionally substituted heterocyclic ring
systems, and further examples of the 5- and 6-membered ring systems
discussed above can be found in W. Durckheimer et aI., U.S. Pat.
No. 4,278,793.
An optionally preferred group of "heterocyclics" include;
1,3-thiazol-2-yl, 4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,
4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,
1,2,4-thiadiazol-5-yl, 3-methyl-1,2,4-thiadiazol-5-yl,
1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl,
2-hydroxy-1,3,4-triazol-5-yl, 2-carboxy-4-methyl-1,3,4-triazol-5-yl
sodium salt, 2-carboxy-4-methyl-1,3,4-triazol-5-yl,
1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl,
2-methyl-1,3,4-oxadiazol-5-yl,
2-(hydroxymethyl)-1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl,
1,3,4-thiadiazol-5-yl, 2-thiol-1,3,4-thiadiazol-5-yl,
2-(methylthio)-1,3,4-thiadiazol-5-yl,
2-amino-1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl,
1-methyl-1H-tetrazol-5-yl,
1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,
1-(carboxymethyl)-1H-tetrazol-5-yl, 1-(carboxymethyl)-
1H-tetrazol-5-yl sodium salt, 1-(methylsulfonic
acid)-1H-tetrazol-5-yl, 1-(methylsulfonic acid)-1H-tetrazol-5-yl
sodium salt, 2-methyl-1H-tetrazol-5-yl, 1,2,3-triazol-5-yl,
1-methyl-1,2,3-triazol-5-yl, 2-methyl-1,2,3-triazol5-yl,
4-methyl-1,2,3-triazol-5-yl, pyrid-2-yl N-oxide,
6-methoxy-2-(n-oxide)-pyridaz-3-yl, 6-hydroxypyridaz-3-yl,
1-methylpyrid-2-yl, 1-methylpyrid-4-yl, 2-hydroxypyrimid-4-yl,
1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,
1,4,5,6-tetrahydro-4-(formylmethyl)-5,6-dioxo-as-triazin-3-yl,
2,5-dihydro-5-oxo-6-hydroxy-astriazin-3-yl,
2,5-dihydro-5-oxo-6-hydroxy-as-triazin-3-yl sodium salt,
2,5-dihydro-5-oxo-6-hydroxy-2-methyl-astriazin-3-yl sodium salt,
2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,
2,5-dihydro-5-oxo-6-methoxy-2-methyl-as-triazin-3-yl,
2,5-dihydro-5-oxo-as-triazin-3-yl,
2,5-dihydro-5-oxo-2-methyl-as-triazin-3-yl,
2,5-dihydro-5-oxo-2,6-dimethyl-as-triazin-3-yl, tetrazolo[1,5-b
]pyridazin-6-yl and 8-aminotetrazolo[ 1,5-b]-pyridazin-6-yl.
An alternative group of "heterocyclics" includes;
4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,
4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,
1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl, 1H-tetrazol-5-yl,
1-methyl-1H-tetrazol-5-yl,
1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,
1-(carboxymethyl)-1H-tetrazol-5-yl,
1-(carboxymethyl)-1H-tetrazol-5-yl sodium salt, 1-(methylsulfonic
acid)-1H-tetrazol-5-yl, 1-(methylsulfonic acid)-1H-tetrazol-5-yl
sodium salt, 1,2,3-triazol-5-yl,
1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,
1,4,5,6-tetrahydro-4-(2-formylmethyl)-5,6-dioxo-as-triazin-3-yl,
2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl sodium salt,
2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,
tetrazolo[1,5-b]pyridazin-6-yl, and
8-aminotetrazolo[1,5-b]pyridazin-6-yl.
The terms "heteroaryl group" or "heteroaryl" are used
interchangeably herein and refer to any mono-, bi-, or tricyclic
aromatic rings having the number of ring atoms designated where at
least one ring is a 5-, 6- or 7-membered hydrocarbon ring
containing from one to four heteroatoms selected from nitrogen,
oxygen, and sulfur, preferably at least one heteroatom is nitrogen.
The aryl portion of the term "heteroaryl" refers to aromaticity, a
term known to those skilled in the art and defined in greater
detail in Advanced Organic Chemistry J. March, 3.sup.rd ed., pages
37-69, John Wiley & Sons, New York (1985).
Each substituent or term used in any formula or expression herein,
e.g., T.sup.1, T.sup.2, W, R.sup.n, R.sup.n', Z, Y.sup.n, Ar,
A.sup.n, X, V, C.sub.1 -C.sub.6 alkyl, etc. when it appears more
than once, is independent of its definition elsewhere in that or
any other formula or structure.
"Pharmaceutically acceptable salts" include both acid and base
addition salts.
"Pharmaceutically acceptable acid addition salt" refers to those
salts which retain the biological effectiveness and properties of
the free bases and which are not biologically or otherwise
undesirable, formed with inorganic acids such as hydrochloric acid,
hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and
the like, and organic acids such as acetic acid, propionic acid,
glycolic acid, pyruvic acid, oxalic acid, maleic acid, maloneic
acid, succinic acid, fumaric acid, tartaric acid, citric acid,
benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,
ethanesulfonic acid, p-toluenesulfonic acid, salicyclic acid and
the like.
"Pharmaceutically acceptable base addition salts" include those
derived from inorganic bases such as sodium, potassium, lithium,
ammonium, calcium, magnesium, iron, zinc, copper, manganese,
aluminum salts and the like. Particularly preferred are the
ammonium, potassium, sodium, calcium and magnesium salts. Salts
derived from pharmaceutically acceptable organic nontoxic bases
includes salts of primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines and basic ion exchange resins, such as
isopropylamine, trimethylamine, diethylamine, triethylamine,
tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, ethylenediamine, glucosamine,
methylglucamine, theobromine, purines, piperizine, piperidine,
N-ethylpiperidine, polyamine resins and the like. Particularly
preferred organic non-toxic bases are isopropylamine, diethylamine,
ethanolamine, trimethamine, dicyclohexylamine, choline, and
caffeine.
The term "prodrug" as used here means a pharmacologically inactive
derivative of a parent drug molecule that requires
biotransformation, either spontaneous or enzymatic, within the
organism to release the active drug.
B. Utility
The present invention is the result of the unexpected discovery
that substituted benzodiazepines and analogs thereof defined by
formulae A, and I-VIII inhibit farnesyl:protein transferase and the
farnsylation of p21.sup.ras. Accordingly, pharmaceutical
compositions containing the compounds of structural formula A
inhibit farnesylation of the oncogene protein ras and are useful as
pharmaceutical agents for mammals, especially humans, for the
treatment of diseases where inhibition of farnesylation is
indicated. In one embodiment of the invention inhibition of
farnesylation is indicated for neoplastic and proliferative
diseases including but not limited to, colorectal carcinoma,
exocrine pancreatic carcinoma, and myeloid leukemias. In an
alternative embodiment, inhibition of farnesylation is contemplated
to be useful for proliferating skin diseases where ras positive
cells are found in the differentiated layer, including but not
limited to, psoriasis vulgaris, lichen planus, verruca vulgaris,
verruca plana juvenills, and seborrheic keratosis. Other ras
positive diseases contemplated to be usefully treated with the
inhibitors of this invention include; neurofibromatosis, rheumatoid
arthritis, human papilloma viral infection, Kapoli's sarcoma,
scleroderma and Aleution disease viral infection.
The present invention is also useful in a method directed to
treating fungal infections in an organism in need of such
treatment, which method comprises administering a non toxic (to the
organism) therapeutically effective amount of compounds represented
by structural formulae A and I-IX. In one embodiment, the fungaily
infected orgamisms are animal, preferably mammal, most preferably
human, especially immunologically compromised individuals. In an
alternative embodiment the organisms are plants infected with or
succeptable to blight, rust, and mildew (especially fusarium wilt).
Optionally, nonliving material such as soil may be usefully treated
with the instant compounds to prevent fungal infection of an
organism.
Finally, the instant compounds may be usefully employed as metal
ion and metalloprotein chelators.
C. Preferred Embodiments
One preferred embodiment of the invention comprises a compound
capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula I' ##STR23## where the substituents R, and
R' are hydrogen or perfluro-lower alkyl, R.sup.4 and R.sup.4' are
hydrogen or lower alkyl, and R.sup.7, W, R.sup.24, and R.sup.25 are
selected according to Table I'.
TABLE I'
__________________________________________________________________________
R.sup.7 W R.sup.24 R.sup.25
__________________________________________________________________________
##STR24## CH.sub.3 ##STR25## H ##STR26## CH.sub.3 ##STR27## H
##STR28## CH.sub.3 ##STR29## H ##STR30## CH.sub.3 ##STR31## H
##STR32## CH.sub.3 ##STR33## H ##STR34## CH.sub.3 ##STR35## H
##STR36## CH.sub.3 ##STR37## H ##STR38## CH.sub.3 ##STR39## H
##STR40## CH.sub.3 ##STR41## H ##STR42## CH.sub.3 ##STR43## H
##STR44## CH.sub.3 ##STR45## H ##STR46## CH.sub.3 ##STR47## H
##STR48## CH.sub.3 ##STR49## H ##STR50## CH.sub.3 ##STR51## H
##STR52## CH.sub.3 ##STR53## H ##STR54## CH.sub.3 ##STR55## H
##STR56## CH.sub.3 ##STR57## H ##STR58## CH.sub.3 ##STR59## H
##STR60## CH.sub.3 ##STR61## H ##STR62## CH.sub.3 ##STR63## H
##STR64## CH.sub.3 ##STR65## H ##STR66## CH.sub.3 ##STR67## H
##STR68## CH.sub.3 ##STR69## H ##STR70## CH.sub.3 ##STR71## H
##STR72## CH.sub.3 ##STR73## H ##STR74## CH.sub.3 ##STR75## H
##STR76## CH.sub.3 ##STR77## H ##STR78## CH.sub.3 ##STR79## H
##STR80## CH.sub.3 ##STR81## H ##STR82## CH.sub.3 ##STR83## H
##STR84## CH.sub.3 ##STR85## H ##STR86## CH.sub.3 ##STR87## H
##STR88## CH.sub.3 ##STR89## H ##STR90## CH.sub.3 ##STR91## H
##STR92## CH.sub.3 ##STR93## H ##STR94## CH.sub.3 ##STR95## H
##STR96## CH.sub.3 ##STR97## H ##STR98## CH.sub.3 ##STR99## H
##STR100## CH.sub.3 ##STR101## H ##STR102## CH.sub.3 ##STR103## H
##STR104## CH.sub.3 ##STR105## H ##STR106## CH.sub.3 ##STR107## H
##STR108## CH.sub.3 ##STR109## H ##STR110## CH.sub.3 ##STR111## H
##STR112## CH.sub.3 ##STR113## H ##STR114## CH.sub.3 ##STR115## H
##STR116## CH.sub.3 ##STR117## H ##STR118## CH.sub.3 ##STR119## H
##STR120## CH.sub.3 ##STR121## H ##STR122## CH.sub.3 ##STR123## H
##STR124## CH.sub.3 ##STR125## H ##STR126## CH.sub.3 ##STR127## H
##STR128## CH.sub.3 ##STR129## H ##STR130## CH.sub.3 ##STR131## H
##STR132## CH.sub.3 ##STR133## H ##STR134## CH.sub.3 ##STR135## H
##STR136## CH.sub.3 ##STR137## H ##STR138## CH.sub.3 ##STR139## H
##STR140## CH.sub.3 ##STR141## H ##STR142## CH.sub.3 ##STR143## H
##STR144## CH.sub.3 ##STR145## H ##STR146## CH.sub.3 ##STR147## H
##STR148## CH.sub.3 ##STR149## H ##STR150## CH.sub.3 ##STR151## H
##STR152## CH.sub.3 ##STR153## H ##STR154## CH.sub.3 ##STR155## H
##STR156## CH.sub.3 ##STR157## H ##STR158## CH.sub.3 ##STR159## H
##STR160## CH.sub.3 ##STR161## H ##STR162## CH.sub.3 ##STR163## H
##STR164## CH.sub.3 ##STR165## H ##STR166## CH.sub.3 ##STR167## H
##STR168## CH.sub.3 ##STR169## H ##STR170## CH.sub.3 ##STR171## H
##STR172## CH.sub.3 ##STR173## H ##STR174## CH.sub.3 ##STR175## H
##STR176## CH.sub.3 ##STR177## H ##STR178## CH.sub.3 ##STR179## H
##STR180## CH.sub.3 ##STR181## H ##STR182## CH.sub.3 ##STR183## H
##STR184## CH.sub.3 ##STR185## H ##STR186## CH.sub.3 ##STR187##
H ##STR188## CH.sub.3 ##STR189## H ##STR190## CH.sub.3 ##STR191## H
##STR192## CH.sub.3 ##STR193## H ##STR194## CH.sub.3 ##STR195## H
##STR196## CH.sub.3 ##STR197## H ##STR198## CH.sub.3 ##STR199## H
##STR200## CH.sub.3 ##STR201## H ##STR202## CH.sub.3 ##STR203## H
##STR204## CH.sub.3 ##STR205## H ##STR206## CH.sub.3 ##STR207## H
##STR208## CH.sub.3 ##STR209## H ##STR210## CH.sub.3 ##STR211## H
##STR212## CH.sub.3 ##STR213## H ##STR214## CH.sub.3 ##STR215## H
##STR216## CH.sub.3 ##STR217## H ##STR218## CH.sub.3 ##STR219## H
##STR220## CH.sub.3 ##STR221## H ##STR222## CH.sub.3 ##STR223## H
##STR224## CH.sub.3 ##STR225## H ##STR226## CH.sub.3 ##STR227## H
##STR228## CH.sub.3 ##STR229## H ##STR230## CH.sub.3 ##STR231## H
##STR232## CH.sub.3 ##STR233## H ##STR234## CH.sub.3 ##STR235## H
##STR236## CH.sub.3 ##STR237## H ##STR238## CH.sub.3 ##STR239## H
##STR240## CH.sub.3 ##STR241## H ##STR242## CH.sub.3 ##STR243## H
##STR244## CH.sub.3 ##STR245## H ##STR246## CH.sub.3 ##STR247## H
##STR248## CH.sub.3 ##STR249## H ##STR250## CH.sub.3 ##STR251## H
##STR252## CH.sub.3 ##STR253## H ##STR254## CH.sub.3 ##STR255## H
##STR256## CH.sub.3 ##STR257## H ##STR258## CH.sub.3 ##STR259## H
##STR260## CH.sub.3 ##STR261## H ##STR262## CH.sub.3 ##STR263## H
##STR264## CH.sub.3 ##STR265## H ##STR266## CH.sub.3 ##STR267## H
##STR268## CH.sub.3 ##STR269## H ##STR270## CH.sub.3 ##STR271## H
##STR272## CH.sub.3 ##STR273## H ##STR274## CH.sub.3 ##STR275## H
##STR276## CH.sub.3 ##STR277## H ##STR278## CH.sub.3 ##STR279## H
##STR280## CH.sub.3 ##STR281## H ##STR282## CH.sub.3 ##STR283## H
##STR284## CH.sub.3 ##STR285## H ##STR286## CH.sub.3 ##STR287## H
##STR288## CH.sub.3 ##STR289## H ##STR290## CH.sub.3 ##STR291## H
##STR292## CH.sub.3 ##STR293## H ##STR294## CH.sub.3 ##STR295## H
##STR296## CH.sub.3 ##STR297## H ##STR298## CH.sub.3 ##STR299## H
##STR300## CH.sub.3 ##STR301## H ##STR302## CH.sub.3 ##STR303## H
##STR304## CH.sub.3 ##STR305## H ##STR306## CH.sub.3 ##STR307## H
##STR308## CH.sub.3 ##STR309## H ##STR310## CH.sub.3 ##STR311## H
##STR312## CH.sub.3 ##STR313## H ##STR314## CH.sub.3 ##STR315## H
##STR316## CH.sub.3 ##STR317## H ##STR318## CH.sub.3 ##STR319## H
##STR320## CH.sub.3 ##STR321## H ##STR322## CH.sub.3 ##STR323## H
##STR324## CH.sub.3 ##STR325## H ##STR326## CH.sub.3 ##STR327## H
##STR328## CH.sub.3 ##STR329## H ##STR330## CH.sub.3 ##STR331## H
##STR332## CH.sub.3 ##STR333## H ##STR334## CH.sub.3 ##STR335## H
##STR336## CH.sub.3 ##STR337## H ##STR338## CH.sub.3 ##STR339## H
##STR340## CH.sub.3 ##STR341## H ##STR342## CH.sub.3 ##STR343## H
##STR344## CH.sub.3 ##STR345## H ##STR346## CH.sub.3 ##STR347## H
##STR348## CH.sub.3 ##STR349## H ##STR350## CH.sub.3 ##STR351## H
##STR352## CH.sub.3 ##STR353## H ##STR354## CH.sub.3
##STR355## H ##STR356## CH.sub.3 ##STR357## H ##STR358## CH.sub.3
##STR359## H ##STR360## CH.sub.3 ##STR361## H ##STR362## CH.sub.3
##STR363## H ##STR364## CH.sub.3 ##STR365## H ##STR366## CH.sub.3
##STR367## H ##STR368## CH.sub.3 ##STR369## H ##STR370## CH.sub.3
##STR371## H ##STR372## CH.sub.3 ##STR373## H ##STR374## CH.sub.3
##STR375## H ##STR376## CH.sub.3 ##STR377## H ##STR378## CH.sub.3
##STR379## H ##STR380## CH.sub.3 ##STR381## H ##STR382## CH.sub.3
##STR383## H ##STR384## CH.sub.3 ##STR385## H ##STR386## CH.sub.3
##STR387## H ##STR388## CH.sub.3 ##STR389## H ##STR390## CH.sub.3
##STR391## H ##STR392## CH.sub. 3 ##STR393## H ##STR394## CH.sub.3
##STR395## H ##STR396## CH.sub.3 ##STR397## H ##STR398## CH.sub.3
##STR399## H ##STR400## CH.sub.3 ##STR401## H ##STR402## CH.sub.3
##STR403## H ##STR404## CH.sub.3 ##STR405## H ##STR406## CH.sub.3
##STR407## H ##STR408## CH.sub.3 ##STR409## H ##STR410## CH.sub.3
##STR411## H ##STR412## CH.sub.3 ##STR413## H ##STR414## CH.sub.3
##STR415## H ##STR416## CH.sub.3 ##STR417## H ##STR418## CH.sub.3
##STR419## H ##STR420## CH.sub.3 ##STR421## H ##STR422## CH.sub.3
##STR423## H ##STR424## CH.sub.3 ##STR425## H ##STR426## CH.sub.3
##STR427## H ##STR428## CH.sub.3 ##STR429## H ##STR430## CH.sub.3
##STR431## H ##STR432## CH.sub.3 ##STR433## H ##STR434## CH.sub.3
##STR435## H ##STR436## CH.sub.3 ##STR437## H ##STR438## CH.sub.3
##STR439## H ##STR440## CH.sub.3 ##STR441## H ##STR442## CH.sub.3
##STR443## H ##STR444## CH.sub.3 ##STR445## H ##STR446## CH.sub.3
##STR447## H ##STR448## CH.sub.3 ##STR449## H ##STR450## CH.sub.3
##STR451## H ##STR452## CH.sub.3 ##STR453## H ##STR454## CH.sub.3
##STR455## H ##STR456## CH.sub.3 ##STR457## H ##STR458## CH.sub.3
##STR459## H ##STR460## CH.sub.3 ##STR461## H ##STR462## CH.sub.3
##STR463## H ##STR464## CH.sub.3 ##STR465## H ##STR466## CH.sub.3
##STR467## H ##STR468## CH.sub.3 ##STR469## H ##STR470## CH.sub.3
##STR471## H ##STR472## CH.sub.3 ##STR473## H ##STR474## CH.sub.3
##STR475## H ##STR476## CH.sub.3 ##STR477## H ##STR478## CH.sub.3
##STR479## H ##STR480## CH.sub.3 ##STR481## H ##STR482## CH.sub.3
##STR483## H ##STR484## CH.sub.3 ##STR485## H ##STR486## CH.sub.3
##STR487## H ##STR488## CH.sub.3 ##STR489## H ##STR490## CH.sub.3
##STR491## H ##STR492## CH.sub.3 ##STR493## H ##STR494## CH.sub.3
##STR495## H ##STR496## CH.sub.3 ##STR497## H ##STR498## CH.sub.3
##STR499## H ##STR500## CH.sub.3 ##STR501## H ##STR502## CH.sub.3
##STR503## H ##STR504## CH.sub.3 ##STR505## H ##STR506## CH.sub.3
##STR507## H ##STR508## CH.sub.3 ##STR509## H ##STR510## CH.sub.3
##STR511## H ##STR512## CH.sub.3 ##STR513## H ##STR514## CH.sub.3
##STR515## H ##STR516## CH.sub.3 ##STR517## H ##STR518## CH.sub.3
##STR519## H ##STR520## CH.sub.3 ##STR521## H
##STR522## CH.sub.3 ##STR523## H ##STR524## CH.sub.3 ##STR525## H
##STR526## CH.sub.3 ##STR527## H ##STR528## CH.sub.3 ##STR529## H
##STR530## CH.sub.3 ##STR531## H ##STR532## CH.sub.3 ##STR533## H
##STR534## CH.sub.3 ##STR535## H ##STR536## CH.sub.3 ##STR537## H
##STR538## CH.sub.3 ##STR539## H ##STR540## CH.sub.3 ##STR541## H
##STR542## CH.sub.3 ##STR543## H ##STR544## CH.sub.3 ##STR545## H
##STR546## CH.sub.3 ##STR547## H ##STR548## CH.sub.3 ##STR549## H
##STR550## CH.sub.3 ##STR551## H ##STR552## CH.sub.3 ##STR553## H
##STR554## CH.sub.3 ##STR555## H ##STR556## CH.sub.3 ##STR557## H
##STR558## CH.sub.3 ##STR559## H ##STR560## CH.sub.3 ##STR561## H
##STR562## CH.sub.3 ##STR563## H ##STR564## CH.sub.3 ##STR565## H
##STR566## CH.sub.3 ##STR567## H ##STR568## CH.sub.3 ##STR569## H
##STR570## CH.sub.3 ##STR571## H ##STR572## CH.sub.3 ##STR573## H
##STR574## CH.sub.3 ##STR575## H ##STR576## CH.sub.3 ##STR577## H
##STR578## CH.sub.3 ##STR579## H ##STR580## CH.sub.3 ##STR581## H
##STR582## CH.sub.3 ##STR583## H ##STR584## CH.sub.3 ##STR585## H
##STR586## CH.sub.3 ##STR587## H ##STR588## CH.sub.3
##STR589## H ##STR590## CH.sub.3 ##STR591## H ##STR592## CH.sub.3
##STR593## H ##STR594## CH.sub.3 ##STR595## H ##STR596## CH.sub.3
##STR597## H ##STR598## CH.sub.3 ##STR599## H ##STR600## CH.sub.3
##STR601## H ##STR602## CH.sub.3 ##STR603## H ##STR604## CH.sub.3
##STR605## H ##STR606## CH.sub.3 ##STR607## H ##STR608## CH.sub.3
##STR609## H ##STR610## CH.sub.3 ##STR611## H ##STR612## CH.sub.3
##STR613## H ##STR614## CH.sub.3 ##STR615## H ##STR616## CH.sub.3
##STR617## H ##STR618## CH.sub.3 ##STR619## H ##STR620## CH.sub.3
##STR621## H ##STR622## CH.sub.3 ##STR623## H ##STR624## CH.sub.3
##STR625## H ##STR626## CH.sub.3 ##STR627## H ##STR628## CH.sub.3
##STR629## H ##STR630## CH.sub. 3 ##STR631## H ##STR632## CH.sub.3
##STR633## H ##STR634## CH.sub.3 ##STR635## H ##STR636## CH.sub.3
##STR637## H ##STR638## CH.sub.3 ##STR639## H ##STR640## CH.sub.3
##STR641## H ##STR642## CH.sub.3 ##STR643## H ##STR644## CH.sub.3
##STR645## H ##STR646## CH.sub.3 ##STR647## H ##STR648## CH.sub.3
##STR649## H ##STR650## CH.sub.3 ##STR651## H ##STR652## CH.sub.3
##STR653## H ##STR654## CH.sub.3 ##STR655## H ##STR656## CH.sub.3
##STR657## H ##STR658## CH.sub.3 ##STR659## H ##STR660## CH.sub.3
##STR661## H ##STR662## CH.sub.3 ##STR663## H ##STR664## CH.sub.3
##STR665## H ##STR666## CH.sub.3 ##STR667## H ##STR668## CH.sub.3
##STR669## H ##STR670## CH.sub.3 ##STR671## H ##STR672## CH.sub.3
##STR673## H ##STR674## CH.sub.3 ##STR675## H ##STR676## CH.sub.3
##STR677## H ##STR678## CH.sub.3 ##STR679## H ##STR680## CH.sub.3
##STR681## H ##STR682## CH.sub.3 ##STR683##
__________________________________________________________________________
An alternate preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula Ia'. ##STR684## where the substituents R,
and R' are as defined above, R.sup.4 and R.sup.4' are hydrogen or
lower alkyl, and R.sup.7', R.sup.8, R.sup.24, and R.sup.25 are
selected according to Table Ia'.
TABLE Ia'
__________________________________________________________________________
R.sup.7 ' R.sup.8 R.sup.24 R.sup.25
__________________________________________________________________________
##STR685## CH.sub.3 ##STR686## H ##STR687## CH.sub.3 ##STR688## H
##STR689## CH.sub.3 ##STR690## H ##STR691## CH.sub.3 ##STR692## H
##STR693## CH.sub.3 ##STR694## H ##STR695## CH.sub.3 ##STR696## H
##STR697## CH.sub.3 ##STR698## H ##STR699## CH.sub.3 ##STR700## H
##STR701## CH.sub.3 ##STR702## H ##STR703## CH.sub.3 ##STR704## H
##STR705## CH.sub.3 ##STR706## H ##STR707## CH.sub.3 ##STR708## H
##STR709## CH.sub.3 ##STR710## H ##STR711## CH.sub.3 ##STR712## H
##STR713## CH.sub.3 ##STR714## H ##STR715## CH.sub.3 ##STR716## H
##STR717## CH.sub.3 ##STR718## H ##STR719## CH.sub.3 ##STR720## H
##STR721## CH.sub.3 ##STR722## H ##STR723## CH.sub.3 ##STR724## H
##STR725## CH.sub.3 ##STR726## H ##STR727## CH.sub.3 ##STR728## H
##STR729## CH.sub.3 ##STR730## H ##STR731## CH.sub.3 ##STR732## H
##STR733## CH.sub.3 ##STR734## H ##STR735## CH.sub.3 ##STR736## H
##STR737## CH.sub.3 ##STR738## H ##STR739## CH.sub.3 ##STR740## H
##STR741## CH.sub.3 ##STR742## H ##STR743## CH.sub.3 ##STR744## H
##STR745## CH.sub.3 ##STR746## H ##STR747## CH.sub.3 ##STR748## H
##STR749## CH.sub.3 ##STR750## H ##STR751## CH.sub.3 ##STR752## H
##STR753## CH.sub.3 ##STR754## H ##STR755## CH.sub.3 ##STR756## H
##STR757## CH.sub.3 ##STR758## H ##STR759## CH.sub.3 ##STR760## H
##STR761## CH.sub.3 ##STR762## H ##STR763## CH.sub.3 ##STR764## H
##STR765## CH.sub.3 ##STR766## H ##STR767## CH.sub.3 ##STR768## H
##STR769## CH.sub.3 ##STR770## H ##STR771## CH.sub.3 ##STR772## H
##STR773## CH.sub.3 ##STR774## H ##STR775## CH.sub.3 ##STR776## H
##STR777## CH.sub.3 ##STR778## H ##STR779## CH.sub.3 ##STR780## H
##STR781## C.sub.2 H.sub.5 ##STR782## H ##STR783## C.sub.2 H.sub.5
##STR784## H ##STR785## C.sub.2 H.sub.5 ##STR786## H ##STR787##
C.sub.2 H.sub.5 ##STR788## H ##STR789## C.sub.2 H.sub.5 ##STR790##
H ##STR791## C.sub.2 H.sub.5 ##STR792## H ##STR793## C.sub.2
H.sub.5 ##STR794## H ##STR795## C.sub.2 H.sub.5 ##STR796## H
##STR797## C.sub.2 H.sub.5 ##STR798## H ##STR799## C.sub.2 H.sub.5
##STR800## H ##STR801## C.sub.2 H.sub.5 ##STR802## H ##STR803##
C.sub.2 H.sub.5 ##STR804## H ##STR805## C.sub.2 H.sub.5 ##STR806##
H ##STR807## C.sub.2 H.sub.5 ##STR808## H ##STR809## C.sub.2
H.sub.5 ##STR810## H ##STR811## C.sub.2 H.sub.5 ##STR812## H
##STR813## CH.sub.3 ##STR814## H ##STR815## CH.sub.3 ##STR816## H
##STR817## CH.sub.3 ##STR818## H ##STR819## CH.sub.3 ##STR820## H
##STR821## CH.sub.3 ##STR822## H ##STR823## CH.sub.3 ##STR824## H
##STR825## CH.sub.3 ##STR826## H ##STR827## CH.sub.3 ##STR828## H
##STR829## CH.sub.3 ##STR830## H ##STR831## CH.sub.3 ##STR832## H
##STR833## CH.sub.3 ##STR834## H ##STR835## CH.sub.3 ##STR836## H
##STR837## CH.sub. 3 ##STR838## H ##STR839## CH.sub.3 ##STR840## H
##STR841## CH.sub.3 ##STR842## H ##STR843## CH.sub.3 ##STR844## H
##STR845## ##STR846## ##STR847## H ##STR848## ##STR849##
##STR850## H ##STR851## ##STR852## ##STR853## H ##STR854##
##STR855## ##STR856## H ##STR857## ##STR858## ##STR859## H
##STR860## ##STR861## ##STR862## H ##STR863## ##STR864## ##STR865##
H ##STR866## ##STR867## ##STR868## H ##STR869## ##STR870##
##STR871## H ##STR872## ##STR873## ##STR874## H ##STR875##
##STR876## ##STR877## H ##STR878## ##STR879## ##STR880## H
##STR881## ##STR882## ##STR883## H ##STR884## ##STR885## ##STR886##
H ##STR887## ##STR888## ##STR889## H ##STR890## ##STR891##
##STR892## CH.sub.3 ##STR893## CH.sub.3 ##STR894## CH.sub.3
##STR895## CH.sub.3 ##STR896## CH.sub.3 ##STR897## CH.sub.3
##STR898## CH.sub.3 ##STR899## CH.sub.3 ##STR900## CH.sub.3
##STR901## CH.sub.3 ##STR902## CH.sub.3 ##STR903## CH.sub.3
##STR904## CH.sub.3 ##STR905## CH.sub.3 ##STR906## CH.sub.3
##STR907## CH.sub.3 ##STR908## CH.sub.3 ##STR909## CH.sub.3
##STR910## CH.sub.3 ##STR911## CH.sub.3 ##STR912## CH.sub.3
##STR913## CH.sub.3 ##STR914## CH.sub.3 ##STR915## CH.sub.3
##STR916## CH.sub.3 ##STR917## CH.sub.3 ##STR918## CH.sub.3
##STR919## CH.sub.3 ##STR920## CH.sub.3 ##STR921## CH.sub.3
##STR922## CH.sub.3 ##STR923## CH.sub.3 ##STR924## H ##STR925##
CH.sub.3 ##STR926## H ##STR927## CH.sub.3 ##STR928## H ##STR929##
CH.sub.3 ##STR930## H ##STR931## CH.sub.3 ##STR932## H ##STR933##
CH.sub.3 ##STR934## H ##STR935## CH.sub.3 ##STR936## H ##STR937##
CH.sub.3 ##STR938## H ##STR939## CH.sub.3 ##STR940## H ##STR941##
CH.sub.3 ##STR942## H ##STR943## CH.sub.3 ##STR944## H ##STR945##
CH.sub.3 ##STR946## H ##STR947## CH.sub.3 ##STR948## H ##STR949##
CH.sub.3 ##STR950## H ##STR951## CH.sub.3 ##STR952## H ##STR953##
CH.sub.3 ##STR954## H ##STR955## CH.sub.3 ##STR956## H ##STR957##
C.sub.4 H.sub.9 ##STR958## H ##STR959## C.sub.4 H.sub.9 ##STR960##
H ##STR961## C.sub.4 H.sub.9 ##STR962## H ##STR963## C.sub.4
H.sub.9 ##STR964## H ##STR965## C.sub.4 H.sub.9 ##STR966## H
##STR967## C.sub.4 H.sub.9 ##STR968## H ##STR969## C.sub.4 H.sub.9
##STR970## H ##STR971## C.sub.4 H.sub.9 ##STR972## H ##STR973##
C.sub.4 H.sub.9 ##STR974## H ##STR975## C.sub.4 H.sub.9 ##STR976##
H ##STR977## C.sub.4 H.sub.9 ##STR978## H ##STR979## C.sub.4
H.sub.9 ##STR980## H ##STR981## C.sub.4 H.sub.9 ##STR982## H
##STR983## C.sub.4 H.sub.9 ##STR984## H ##STR985## C.sub.4 H.sub.9
##STR986## H ##STR987## C.sub.4 H.sub.9 ##STR988## H ##STR989##
CH.sub.3 ##STR990## H ##STR991## CH.sub.3 ##STR992## H ##STR993##
CH.sub.3 ##STR994## H ##STR995## CH.sub.3 ##STR996## H ##STR997##
CH.sub.3 ##STR998## H ##STR999## CH.sub.3 ##STR1000## H ##STR1001##
CH.sub.3 ##STR1002## H ##STR1003## CH.sub.3 ##STR1004## H
##STR1005## CH.sub.3 ##STR1006## H ##STR1007## CH.sub.3 ##STR1008##
H ##STR1009## CH.sub.3 ##STR1010## H ##STR1011## CH.sub.3
##STR1012## H ##STR1013## CH.sub.3 ##STR1014## H ##STR1015##
CH.sub.3 ##STR1016## H ##STR1017## CH.sub.3 ##STR1018## H
##STR1019## CH.sub.3 ##STR1020## H ##STR1021## CH.sub.3
##STR1022## H ##STR1023## CH.sub.3 ##STR1024## H ##STR1025##
CH.sub.3 ##STR1026## H ##STR1027## CH.sub.3 ##STR1028## H
##STR1029## CH.sub.3 ##STR1030## H ##STR1031## CH.sub.3 ##STR1032##
H ##STR1033## CH.sub.3 ##STR1034## H ##STR1035## CH.sub.3
##STR1036## H ##STR1037## CH.sub.3 ##STR1038## H ##STR1039##
CH.sub.3 ##STR1040## H ##STR1041## CH.sub.3 ##STR1042## H
##STR1043## CH.sub.3 ##STR1044## H ##STR1045## CH.sub.3 ##STR1046##
H ##STR1047## CH.sub.3 ##STR1048## H ##STR1049## CH.sub.3
##STR1050## H ##STR1051## CH.sub.3 ##STR1052## H ##STR1053##
CH.sub.3 ##STR1054## H ##STR1055## CH.sub.3 ##STR1056## H
##STR1057## CH.sub.3 ##STR1058## H ##STR1059## CH.sub.3 ##STR1060##
H ##STR1061## CH.sub.3 ##STR1062## H ##STR1063## CH.sub.3
##STR1064## H ##STR1065## CH.sub.3 ##STR1066## H ##STR1067##
CH.sub.3 ##STR1068## H ##STR1069## CH.sub.3 ##STR1070## H
##STR1071## CH.sub.3 ##STR1072## H ##STR1073## CH.sub.3 ##STR1074##
H ##STR1075## CH.sub.3 ##STR1076## H ##STR1077## CH.sub.3
##STR1078## H ##STR1079## CH.sub.3 ##STR1080## H ##STR1081##
CH.sub.3 ##STR1082## H ##STR1083## CH.sub.3 ##STR1084## H
##STR1085## CH.sub.3 ##STR1086## H ##STR1087## CH.sub.3 ##STR1088##
H ##STR1089## CH.sub.3 ##STR1090## H ##STR1091## CH.sub.3
##STR1092## H ##STR1093## CH.sub.3 ##STR1094## H ##STR1095##
CH.sub.3 ##STR1096## H ##STR1097## CH.sub.3 ##STR1098## H
##STR1099## CH.sub.3 ##STR1100## H ##STR1101## CH.sub.3 ##STR1102##
H ##STR1103## CH.sub.3 ##STR1104## H ##STR1105## CH.sub.3
##STR1106## H ##STR1107## CH.sub.3 ##STR1108## H ##STR1109##
CH.sub.3 ##STR1110## H ##STR1111## CH.sub.3 ##STR1112## H
##STR1113## CH.sub.3 ##STR1114## H ##STR1115## CH.sub.3 ##STR1116##
H ##STR1117## CH.sub.3 ##STR1118## H ##STR1119## CH.sub.3
##STR1120## H ##STR1121## CH.sub.3 ##STR1122## H ##STR1123##
CH.sub.3 ##STR1124## H ##STR1125## CH.sub.3 ##STR1126## H
##STR1127## CH.sub.3 ##STR1128## H ##STR1129## CH.sub.3 ##STR1130##
H ##STR1131## CH.sub.3 ##STR1132## H ##STR1133## CH.sub.3
##STR1134## H ##STR1135## CH.sub.3 ##STR1136## H ##STR1137##
CH.sub.3 ##STR1138## H ##STR1139## CH.sub.3 ##STR1140## H
##STR1141## CH.sub.3 ##STR1142## H ##STR1143## CH.sub.3 ##STR1144##
H ##STR1145## CH.sub.3 ##STR1146## H ##STR1147## CH.sub.3
##STR1148## H ##STR1149## CH.sub.3 ##STR1150## H ##STR1151##
CH.sub.3 ##STR1152## H ##STR1153## CH.sub.3 ##STR1154## H
##STR1155## CH.sub.3 ##STR1156## H ##STR1157## CH.sub.3 ##STR1158##
H ##STR1159## CH.sub.3 ##STR1160## H ##STR1161## CH.sub. 3
##STR1162## H ##STR1163## CH.sub.3 ##STR1164## H ##STR1165##
CH.sub.3 ##STR1166## H ##STR1167## CH.sub.3 ##STR1168## H
##STR1169## CH.sub.3 ##STR1170## H ##STR1171## CH.sub.3 ##STR1172##
H ##STR1173## CH.sub.3 ##STR1174## H ##STR1175## CH.sub.3
##STR1176## H ##STR1177## CH.sub.3 ##STR1178## H ##STR1179##
CH.sub.3 ##STR1180## H ##STR1181## CH.sub.3 ##STR1182## H
##STR1183## CH.sub.3 ##STR1184## H ##STR1185## CH.sub.3 ##STR1186##
H ##STR1187## CH.sub.3 ##STR1188## H
##STR1189## CH.sub.3 ##STR1190## H ##STR1191## CH.sub.3 ##STR1192##
H ##STR1193## CH.sub.3 ##STR1194## H ##STR1195## CH.sub.3
##STR1196## H ##STR1197## CH.sub.3 ##STR1198## H ##STR1199##
CH.sub.3 ##STR1200## H ##STR1201## CH.sub.3 ##STR1202## H
##STR1203## CH.sub.3 ##STR1204## H ##STR1205## CH.sub.3 ##STR1206##
H ##STR1207## CH.sub.3 ##STR1208## H ##STR1209## CH.sub.3
##STR1210## H ##STR1211## CH.sub.3 ##STR1212## H ##STR1213##
CH.sub.3 ##STR1214## H ##STR1215## CH.sub.3 ##STR1216## H
##STR1217## CH.sub.3 ##STR1218## H ##STR1219## CH.sub.3 ##STR1220##
H ##STR1221## CH.sub.3 ##STR1222## H ##STR1223## CH.sub.3
##STR1224## H ##STR1225## CH.sub.3 ##STR1226## H ##STR1227##
CH.sub.3 ##STR1228## H ##STR1229## CH.sub.3 ##STR1230## H
##STR1231## CH.sub.3 ##STR1232## H ##STR1233## CH.sub.3 ##STR1234##
H ##STR1235## CH.sub.3 ##STR1236## H ##STR1237## CH.sub.3
##STR1238## H ##STR1239## CH.sub.3 ##STR1240## H ##STR1241##
CH.sub.3 ##STR1242## H ##STR1243## CH.sub.3 ##STR1244## H
##STR1245## CH.sub.3 ##STR1246## H ##STR1247## CH.sub.3 ##STR1248##
H ##STR1249## CH.sub.3 ##STR1250## H ##STR1251## CH.sub.3
##STR1252## H ##STR1253## CH.sub.3 ##STR1254## H ##STR1255##
CH.sub.3
##STR1256## H ##STR1257## CH.sub.3 ##STR1258## H ##STR1259##
CH.sub.3 ##STR1260## H ##STR1261## CH.sub.3 ##STR1262## H
##STR1263## CH.sub.3 ##STR1264## H ##STR1265## CH.sub.3 ##STR1266##
H ##STR1267## CH.sub.3 ##STR1268## H ##STR1269## CH.sub.3
##STR1270## H ##STR1271## CH.sub.3 ##STR1272## H ##STR1273##
CH.sub.3 ##STR1274## H ##STR1275## CH.sub.3 ##STR1276## H
##STR1277## CH.sub.3 ##STR1278## H ##STR1279## CH.sub.3 ##STR1280##
H ##STR1281## CH.sub.3 ##STR1282## H ##STR1283## CH.sub.3
##STR1284## H ##STR1285## CH.sub.3 ##STR1286## H ##STR1287##
CH.sub.3 ##STR1288## H ##STR1289## CH.sub.3 ##STR1290## H
##STR1291## CH.sub.3 ##STR1292## H ##STR1293## CH.sub.3 ##STR1294##
H ##STR1295## CH.sub.3 ##STR1296## H ##STR1297## CH.sub.3
##STR1298## H ##STR1299## CH.sub.3 ##STR1300## H ##STR1301##
CH.sub.3 ##STR1302## H ##STR1303## CH.sub.3 ##STR1304## H
##STR1305## CH.sub.3 ##STR1306## H ##STR1307## CH.sub.3 ##STR1308##
H ##STR1309## CH.sub.3 ##STR1310## H ##STR1311## CH.sub.3
##STR1312## H ##STR1313## CH.sub.3 ##STR1314## H ##STR1315##
CH.sub.3 ##STR1316## H ##STR1317## CH.sub.3 ##STR1318## H
##STR1319## CH.sub.3 ##STR1320## H ##STR1321## CH.sub.3 ##STR1322##
H ##STR1323## CH.sub.3 ##STR1324## H ##STR1325## CH.sub.3
##STR1326## H ##STR1327## CH.sub.3 ##STR1328## H ##STR1329##
CH.sub.3 ##STR1330## H ##STR1331## CH.sub.3 ##STR1332## H
##STR1333## CH.sub.3 ##STR1334## H ##STR1335## CH.sub.3 ##STR1336##
H ##STR1337## CH.sub. 3 ##STR1338## H ##STR1339## CH.sub.3
##STR1340## H ##STR1341## CH.sub.3 ##STR1342## H ##STR1343##
CH.sub.3 ##STR1344## H ##STR1345## CH.sub.3 ##STR1346## H
##STR1347## CH.sub.3 ##STR1348## H ##STR1349## CH.sub.3 ##STR1350##
H ##STR1351## CH.sub.3 ##STR1352## H ##STR1353## CH.sub.3
##STR1354## H ##STR1355## CH.sub.3 ##STR1356## H ##STR1357##
CH.sub.3 ##STR1358## H ##STR1359## CH.sub.3 ##STR1360## H
##STR1361## CH.sub.3 ##STR1362## H ##STR1363## CH.sub.3 ##STR1364##
H ##STR1365## CH.sub.3 ##STR1366## H ##STR1367## CH.sub.3
##STR1368## H ##STR1369## CH.sub.3 ##STR1370## H ##STR1371##
CH.sub.3 ##STR1372## H ##STR1373## CH.sub.3 ##STR1374## H
##STR1375## CH.sub.3 ##STR1376## H ##STR1377## CH.sub.3 ##STR1378##
H ##STR1379## CH.sub.3 ##STR1380## H ##STR1381## CH.sub.3
##STR1382## H ##STR1383## CH.sub.3 ##STR1384## H ##STR1385##
CH.sub.3 ##STR1386## H ##STR1387## CH.sub.3 ##STR1388## H
##STR1389## CH.sub.3 ##STR1390## H ##STR1391## CH.sub.3 ##STR1392##
H ##STR1393## CH.sub.3 ##STR1394## H ##STR1395## CH.sub.3
##STR1396## H ##STR1397## CH.sub.3 ##STR1398## H ##STR1399##
CH.sub.3 ##STR1400## H ##STR1401## CH.sub.3 ##STR1402## H
##STR1403## CH.sub.3 ##STR1404##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula II' ##STR1405## where the substituents R and
R' are as defined above, R.sup.4 and R.sup.4' are hydrogen or lower
alkyl, and where W, R.sup.1 and R.sup.2, R.sup.24 and R.sup.25, are
selected according to Table II'.
TABLE II'
__________________________________________________________________________
R.sup.1 and R.sup.2 W R.sup.24 R.sup.25
__________________________________________________________________________
CF.sub.3, H ##STR1406## CH.sub.3 ##STR1407## CF.sub.3, H
##STR1408## CH.sub.3 ##STR1409## CF.sub.3, H ##STR1410## CH.sub.3
##STR1411## CF.sub.3, H ##STR1412## CH.sub.3 ##STR1413## CF.sub.3,
H ##STR1414## CH.sub.3 ##STR1415## CF.sub.3, H ##STR1416## CH.sub.3
##STR1417## CF.sub.3, H ##STR1418## CH.sub.3 ##STR1419## CF.sub.3,
H ##STR1420## CH.sub.3 ##STR1421## CF.sub.3, H ##STR1422## CH.sub.3
##STR1423## CF.sub.3, H ##STR1424## CH.sub.3 ##STR1425## CF.sub.3,
H ##STR1426## CH.sub.3 ##STR1427## CF.sub.3, H ##STR1428## CH.sub.3
##STR1429## CF.sub.3, H ##STR1430## CH.sub.3 ##STR1431## CF.sub.3,
H ##STR1432## CH.sub.3 ##STR1433## CF.sub.3, H ##STR1434## CH.sub.3
##STR1435## CF.sub.3, H ##STR1436## CH.sub.3 ##STR1437## CF.sub.3,
H ##STR1438## CH.sub.3 ##STR1439## CF.sub.3, H ##STR1440## CH.sub.3
##STR1441## CF.sub.3, H ##STR1442## CH.sub.3 ##STR1443## CF.sub.3,
H ##STR1444## CH.sub.3 ##STR1445## CF.sub.3, H ##STR1446## CH.sub.3
##STR1447## CF.sub.3, H ##STR1448## CH.sub.3 ##STR1449## covalent
bond ##STR1450## CH.sub.3 ##STR1451## covalent bond ##STR1452##
CH.sub.3 ##STR1453## covalent bond ##STR1454## CH.sub.3 ##STR1455##
covalent bond ##STR1456## CH.sub.3 ##STR1457## covalent bond
##STR1458## CH.sub.3 ##STR1459## covalent bond ##STR1460## CH.sub.3
##STR1461## covalent bond ##STR1462## CH.sub.3 ##STR1463## covalent
bond ##STR1464## CH.sub.3 ##STR1465## covalent bond ##STR1466##
CH.sub.3 ##STR1467## covalent bond ##STR1468## CH.sub.3 ##STR1469##
covalent bond ##STR1470## CH.sub.3 ##STR1471## ##STR1472##
##STR1473## CH.sub. 3 ##STR1474## ##STR1475## ##STR1476## CH.sub.3
##STR1477## ##STR1478## ##STR1479## CH.sub.3 ##STR1480##
##STR1481## ##STR1482## CH.sub.3 ##STR1483## ##STR1484##
##STR1485## CH.sub.3 ##STR1486## ##STR1487## ##STR1488## CH.sub.3
##STR1489## covalent bond ##STR1490## CH.sub.3 ##STR1491## covalent
bond ##STR1492## CH.sub.3 ##STR1493## covalent bond ##STR1494##
CH.sub.3 ##STR1495## covalent bond ##STR1496## CH.sub.3 ##STR1497##
covalent bond ##STR1498## CH.sub.3 ##STR1499## covalent bond
##STR1500## CH.sub.3 ##STR1501## covalent bond ##STR1502## CH.sub.3
##STR1503## covalent bond ##STR1504## CH.sub.3 ##STR1505## covalent
bond ##STR1506## CH.sub.3 ##STR1507## covalent bond ##STR1508##
CH.sub.3 ##STR1509## covalent bond ##STR1510## CH.sub.3 ##STR1511##
##STR1512## ##STR1513## CH.sub.3 ##STR1514## ##STR1515##
##STR1516## CH.sub.3 ##STR1517## ##STR1518## ##STR1519## CH.sub.3
##STR1520## ##STR1521## ##STR1522## CH.sub.3 ##STR1523##
##STR1524## ##STR1525## CH.sub.3 ##STR1526## ##STR1527##
##STR1528## CH.sub.3 ##STR1529## ##STR1530## ##STR1531## CH.sub.3
##STR1532## ##STR1533## ##STR1534## CH.sub.3 ##STR1535##
##STR1536## ##STR1537## CH.sub.3 ##STR1538## ##STR1539##
##STR1540## CH.sub.3 ##STR1541## ##STR1542## ##STR1543## CH.sub.3
##STR1544## ##STR1545## ##STR1546## CH.sub.3 ##STR1547##
##STR1548## ##STR1549## CH.sub.3 ##STR1550## ##STR1551##
##STR1552## CH.sub.3 ##STR1553## ##STR1554## ##STR1555## CH.sub.3
##STR1556## ##STR1557## ##STR1558## CH.sub.3 ##STR1559##
##STR1560## ##STR1561## CH.sub.3 ##STR1562## ##STR1563##
##STR1564## CH.sub.3 ##STR1565## ##STR1566## ##STR1567## CH.sub.3
##STR1568## ##STR1569## ##STR1570## CH.sub.3 ##STR1571##
##STR1572## ##STR1573## CH.sub.3 ##STR1574## ##STR1575##
##STR1576## CH.sub.3 ##STR1577## ##STR1578## ##STR1579## CH.sub.3
##STR1580## ##STR1581## ##STR1582## CH.sub.3 ##STR1583##
##STR1584## ##STR1585## CH.sub.3 ##STR1586## ##STR1587##
##STR1588## CH.sub.3 ##STR1589## ##STR1590## ##STR1591## CH.sub.3
##STR1592## ##STR1593## ##STR1594## CH.sub.3 ##STR1595##
##STR1596## ##STR1597## CH.sub.3 ##STR1598## ##STR1599##
##STR1600## CH.sub.3 ##STR1601## ##STR1602## ##STR1603## CH.sub.3
##STR1604## ##STR1605## ##STR1606## CH.sub.3
##STR1607## ##STR1608## ##STR1609## CH.sub.3 ##STR1610##
##STR1611## ##STR1612## CH.sub.3 ##STR1613## ##STR1614##
##STR1615## CH.sub.3 ##STR1616## ##STR1617## ##STR1618## CH.sub.3
##STR1619## ##STR1620## ##STR1621## CH.sub.3 ##STR1622##
##STR1623## ##STR1624## CH.sub.3 ##STR1625##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula III' ##STR1626## where the substituents R
and R' are as defined above, R.sup.4 and R.sup.4' are hydrogen or
lower alkyl, and W, R.sup.25 and ##STR1627## are selected according
to Table III'.
TABLE III'
__________________________________________________________________________
##STR1628## ##STR1629## ##STR1630##
__________________________________________________________________________
##STR1631## ##STR1632## ##STR1633## ##STR1634## ##STR1635##
##STR1636## ##STR1637## ##STR1638## ##STR1639## ##STR1640##
##STR1641## ##STR1642## ##STR1643## ##STR1644## ##STR1645##
##STR1646## ##STR1647## ##STR1648## ##STR1649## ##STR1650##
##STR1651## ##STR1652## ##STR1653## ##STR1654## ##STR1655##
##STR1656## ##STR1657## ##STR1658## ##STR1659## ##STR1660##
##STR1661## ##STR1662## ##STR1663## ##STR1664## ##STR1665##
##STR1666## ##STR1667## ##STR1668## ##STR1669## ##STR1670##
##STR1671## ##STR1672## ##STR1673## ##STR1674## ##STR1675##
##STR1676## ##STR1677## ##STR1678## ##STR1679## ##STR1680##
##STR1681## ##STR1682## ##STR1683## ##STR1684## ##STR1685##
##STR1686## ##STR1687## ##STR1688## ##STR1689## ##STR1690##
##STR1691## ##STR1692## ##STR1693## ##STR1694## ##STR1695##
##STR1696## ##STR1697## ##STR1698## ##STR1699## ##STR1700##
##STR1701## ##STR1702## ##STR1703## ##STR1704## ##STR1705##
##STR1706## ##STR1707## ##STR1708## ##STR1709## ##STR1710##
##STR1711## ##STR1712## ##STR1713## ##STR1714## ##STR1715##
##STR1716## ##STR1717## ##STR1718## ##STR1719## ##STR1720##
##STR1721## ##STR1722## ##STR1723## ##STR1724## ##STR1725##
##STR1726## ##STR1727## ##STR1728## ##STR1729## ##STR1730##
##STR1731## ##STR1732## ##STR1733## ##STR1734## ##STR1735##
##STR1736## ##STR1737## ##STR1738## ##STR1739## ##STR1740##
##STR1741## ##STR1742## ##STR1743## ##STR1744## ##STR1745##
##STR1746## ##STR1747## ##STR1748## ##STR1749## ##STR1750##
##STR1751## ##STR1752## ##STR1753## ##STR1754## ##STR1755##
##STR1756## ##STR1757## ##STR1758## ##STR1759## ##STR1760##
##STR1761## ##STR1762## ##STR1763## ##STR1764## ##STR1765##
##STR1766## ##STR1767## ##STR1768## ##STR1769## ##STR1770##
##STR1771## ##STR1772## ##STR1773## ##STR1774## ##STR1775##
##STR1776## ##STR1777## ##STR1778## ##STR1779## ##STR1780##
##STR1781## ##STR1782## ##STR1783## ##STR1784## ##STR1785##
##STR1786## ##STR1787## ##STR1788## ##STR1789## ##STR1790##
##STR1791## ##STR1792## ##STR1793## ##STR1794## ##STR1795##
##STR1796## ##STR1797## ##STR1798## ##STR1799## ##STR1800##
##STR1801## ##STR1802## ##STR1803## ##STR1804## ##STR1805##
##STR1806## ##STR1807## ##STR1808## ##STR1809## ##STR1810##
##STR1811## ##STR1812## ##STR1813## ##STR1814## ##STR1815##
##STR1816## ##STR1817## ##STR1818## ##STR1819## ##STR1820##
##STR1821## ##STR1822## ##STR1823## ##STR1824## ##STR1825##
##STR1826## ##STR1827## ##STR1828## ##STR1829## ##STR1830##
##STR1831## ##STR1832## ##STR1833## ##STR1834## ##STR1835##
##STR1836## ##STR1837## ##STR1838## ##STR1839## ##STR1840##
##STR1841## ##STR1842## ##STR1843## ##STR1844## ##STR1845##
##STR1846## ##STR1847## ##STR1848## ##STR1849## ##STR1850##
##STR1851## ##STR1852## ##STR1853## ##STR1854## ##STR1855##
##STR1856## ##STR1857## ##STR1858## ##STR1859## ##STR1860##
##STR1861## ##STR1862## ##STR1863## ##STR1864## ##STR1865##
##STR1866## ##STR1867## ##STR1868## ##STR1869## ##STR1870##
##STR1871## ##STR1872## ##STR1873## ##STR1874## ##STR1875##
##STR1876## ##STR1877## ##STR1878## ##STR1879## ##STR1880##
##STR1881## ##STR1882## ##STR1883## ##STR1884## ##STR1885##
##STR1886## ##STR1887## ##STR1888## ##STR1889## ##STR1890##
##STR1891## ##STR1892## ##STR1893## ##STR1894##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula IV' ##STR1895## where the substituents R and
R' are as defined above, R.sup.4 and R.sup.4' are hydrogen or lower
alkyl, and W, R.sup.25 and ##STR1896## are selected according to
Table IV'.
TABLE IV'
__________________________________________________________________________
##STR1897## ##STR1898## ##STR1899##
__________________________________________________________________________
##STR1900## ##STR1901## ##STR1902## ##STR1903## ##STR1904##
##STR1905## ##STR1906## ##STR1907## ##STR1908## ##STR1909##
##STR1910## ##STR1911## ##STR1912## ##STR1913## ##STR1914##
##STR1915## ##STR1916## ##STR1917## ##STR1918## ##STR1919##
##STR1920## ##STR1921## ##STR1922## ##STR1923## ##STR1924##
##STR1925## ##STR1926## ##STR1927## ##STR1928## ##STR1929##
##STR1930## ##STR1931## ##STR1932## ##STR1933## ##STR1934##
##STR1935## ##STR1936## ##STR1937## ##STR1938## ##STR1939##
##STR1940## ##STR1941## ##STR1942## ##STR1943## ##STR1944##
##STR1945## ##STR1946## ##STR1947## ##STR1948## ##STR1949##
##STR1950## ##STR1951## ##STR1952## ##STR1953## ##STR1954##
##STR1955## ##STR1956## ##STR1957## ##STR1958## ##STR1959##
##STR1960## ##STR1961## ##STR1962## ##STR1963## ##STR1964##
##STR1965## ##STR1966## ##STR1967## ##STR1968## ##STR1969##
##STR1970## ##STR1971## ##STR1972## ##STR1973## ##STR1974##
##STR1975## ##STR1976## ##STR1977## ##STR1978## ##STR1979##
##STR1980## ##STR1981## ##STR1982## ##STR1983## ##STR1984##
##STR1985## ##STR1986## ##STR1987## ##STR1988## ##STR1989##
##STR1990## ##STR1991## ##STR1992## ##STR1993## ##STR1994##
##STR1995## ##STR1996## ##STR1997## ##STR1998## ##STR1999##
##STR2000## ##STR2001## ##STR2002## ##STR2003## ##STR2004##
##STR2005## ##STR2006## ##STR2007## ##STR2008## ##STR2009##
##STR2010## ##STR2011## ##STR2012## ##STR2013## ##STR2014##
##STR2015## ##STR2016## ##STR2017## ##STR2018## ##STR2019##
##STR2020## ##STR2021## ##STR2022## ##STR2023## ##STR2024##
##STR2025## ##STR2026## ##STR2027## ##STR2028## ##STR2029##
##STR2030## ##STR2031## ##STR2032## ##STR2033## ##STR2034##
##STR2035## ##STR2036## ##STR2037## ##STR2038## ##STR2039##
##STR2040## ##STR2041## ##STR2042## ##STR2043## ##STR2044##
##STR2045## ##STR2046## ##STR2047## ##STR2048## ##STR2049##
##STR2050## ##STR2051## ##STR2052## ##STR2053## ##STR2054##
##STR2055## ##STR2056## ##STR2057## ##STR2058## ##STR2059##
##STR2060## ##STR2061## ##STR2062## ##STR2063## ##STR2064##
##STR2065## ##STR2066## ##STR2067## ##STR2068## ##STR2069##
##STR2070## ##STR2071## ##STR2072## ##STR2073## ##STR2074##
##STR2075## ##STR2076## ##STR2077## ##STR2078## ##STR2079##
##STR2080## ##STR2081## ##STR2082## ##STR2083## ##STR2084##
##STR2085## ##STR2086## ##STR2087## ##STR2088## ##STR2089##
##STR2090## ##STR2091## ##STR2092## ##STR2093## ##STR2094##
##STR2095## ##STR2096## ##STR2097## ##STR2098## ##STR2099##
##STR2100## ##STR2101## ##STR2102## ##STR2103## ##STR2104##
##STR2105## ##STR2106## ##STR2107## ##STR2108## ##STR2109##
##STR2110## ##STR2111## ##STR2112## ##STR2113## ##STR2114##
##STR2115## ##STR2116## ##STR2117## ##STR2118## ##STR2119##
##STR2120## ##STR2121## ##STR2122## ##STR2123## ##STR2124##
##STR2125## ##STR2126## ##STR2127## ##STR2128## ##STR2129##
##STR2130## ##STR2131## ##STR2132## ##STR2133## ##STR2134##
##STR2135## ##STR2136## ##STR2137## ##STR2138## ##STR2139##
##STR2140## ##STR2141## ##STR2142## ##STR2143## ##STR2144##
##STR2145## ##STR2146## ##STR2147## ##STR2148## ##STR2149##
##STR2150## ##STR2151## ##STR2152## ##STR2153## ##STR2154##
##STR2155## ##STR2156## ##STR2157## ##STR2158## ##STR2159##
##STR2160## ##STR2161## ##STR2162## ##STR2163##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula V' ##STR2164## where the substituents R and
R' are halo or perfluro-loweralkyl, R.sup.4 is hydrogen or lower
alkyl, and R.sup.24, R.sup.25 and ##STR2165## are selected
according to Table V'.
TABLE V'
__________________________________________________________________________
##STR2166## ##STR2167## ##STR2168## ##STR2169##
__________________________________________________________________________
##STR2170## ##STR2171## CH.sub.3 ##STR2172## ##STR2173##
##STR2174## CH.sub.3 ##STR2175## ##STR2176## ##STR2177## CH.sub.3
##STR2178## ##STR2179## ##STR2180## CH.sub.3 ##STR2181##
##STR2182## ##STR2183## CH.sub.3 ##STR2184## ##STR2185##
##STR2186## CH.sub.3 ##STR2187## ##STR2188## ##STR2189## CH.sub.3
##STR2190## ##STR2191## ##STR2192## CH.sub.3 ##STR2193##
##STR2194## ##STR2195## CH.sub.3 ##STR2196## ##STR2197##
##STR2198## CH.sub.3 ##STR2199## ##STR2200## ##STR2201## CH.sub.3
##STR2202## ##STR2203## ##STR2204## CH.sub.3 ##STR2205##
##STR2206## ##STR2207## CH.sub.3 ##STR2208## ##STR2209##
##STR2210## CH.sub.3 ##STR2211## ##STR2212## ##STR2213## CH.sub.3
##STR2214## ##STR2215## ##STR2216## CH.sub.3 ##STR2217##
##STR2218## ##STR2219## CH.sub.3 ##STR2220## ##STR2221##
##STR2222## CH.sub.3 ##STR2223## ##STR2224## ##STR2225## CH.sub.3
##STR2226## ##STR2227## ##STR2228## CH.sub.3 ##STR2229##
##STR2230## ##STR2231## CH.sub.3 ##STR2232## ##STR2233##
##STR2234## CH.sub.3 ##STR2235## ##STR2236## ##STR2237## CH.sub.3
##STR2238## ##STR2239## ##STR2240## CH.sub.3 ##STR2241##
##STR2242## ##STR2243## CH.sub.3 ##STR2244## ##STR2245##
##STR2246## CH.sub.3 ##STR2247## ##STR2248## ##STR2249## CH.sub.3
##STR2250## ##STR2251## ##STR2252## CH.sub.3 ##STR2253##
##STR2254## ##STR2255## CH.sub.3 ##STR2256## ##STR2257##
##STR2258## CH.sub.3 ##STR2259## ##STR2260## ##STR2261## CH.sub.3
##STR2262## ##STR2263## ##STR2264## CH.sub.3 ##STR2265##
##STR2266## ##STR2267## CH.sub.3 ##STR2268## ##STR2269##
##STR2270## CH.sub.3 ##STR2271## ##STR2272## ##STR2273## CH.sub.3
##STR2274## ##STR2275## ##STR2276## CH.sub.3 ##STR2277##
##STR2278## ##STR2279## CH.sub.3 ##STR2280## ##STR2281##
##STR2282## CH.sub.3 ##STR2283## ##STR2284## ##STR2285## CH.sub.3
##STR2286## ##STR2287## ##STR2288## CH.sub.3 ##STR2289##
##STR2290## ##STR2291## CH.sub.3 ##STR2292## ##STR2293##
##STR2294## CH.sub.3 ##STR2295## ##STR2296## ##STR2297## CH.sub.3
##STR2298## ##STR2299## ##STR2300## CH.sub.3 ##STR2301##
##STR2302## ##STR2303## CH.sub.3 ##STR2304## ##STR2305##
##STR2306## CH.sub.3 ##STR2307## ##STR2308## ##STR2309## CH.sub.3
##STR2310## ##STR2311## ##STR2312## CH.sub.3 ##STR2313##
##STR2314## ##STR2315## CH.sub.3 ##STR2316## ##STR2317##
##STR2318## CH.sub.3 ##STR2319## ##STR2320## ##STR2321## CH.sub.3
##STR2322## ##STR2323## ##STR2324## CH.sub.3 ##STR2325##
##STR2326## ##STR2327## CH.sub.3 ##STR2328## ##STR2329##
##STR2330## CH.sub.3 ##STR2331## ##STR2332## ##STR2333## CH.sub.3
##STR2334## ##STR2335## ##STR2336## CH.sub.3 ##STR2337##
##STR2338## ##STR2339## CH.sub.3 ##STR2340## ##STR2341##
##STR2342## CH.sub.3 ##STR2343## ##STR2344## ##STR2345## CH.sub.3
##STR2346## ##STR2347## ##STR2348## CH.sub.3 ##STR2349##
##STR2350## ##STR2351## CH.sub.3 ##STR2352## ##STR2353##
##STR2354## CH.sub.3 ##STR2355## ##STR2356## ##STR2357## CH.sub.3
##STR2358## ##STR2359## ##STR2360## CH.sub.3 ##STR2361##
##STR2362## ##STR2363## CH.sub.3 ##STR2364## ##STR2365##
##STR2366## CH.sub.3 ##STR2367## ##STR2368## ##STR2369## CH.sub.3
##STR2370## ##STR2371## ##STR2372## CH.sub.3 ##STR2373##
##STR2374## ##STR2375## CH.sub.3 ##STR2376## ##STR2377##
##STR2378## CH.sub.3 ##STR2379## ##STR2380## ##STR2381## CH.sub.3
##STR2382## ##STR2383## ##STR2384## CH.sub.3 ##STR2385##
##STR2386## ##STR2387## CH.sub.3 ##STR2388## ##STR2389##
##STR2390## CH.sub.3 ##STR2391## ##STR2392## ##STR2393## CH.sub.3
##STR2394## ##STR2395## ##STR2396## CH.sub.3 ##STR2397##
##STR2398## ##STR2399## CH.sub.3 ##STR2400## ##STR2401##
##STR2402## CH.sub.3 ##STR2403## ##STR2404## ##STR2405## CH.sub.3
##STR2406## ##STR2407## ##STR2408## CH.sub.3 ##STR2409##
##STR2410## ##STR2411## CH.sub.3 ##STR2412## ##STR2413##
##STR2414## CH.sub.3 ##STR2415## ##STR2416## ##STR2417## CH.sub.3
##STR2418## ##STR2419## ##STR2420## CH.sub.3 ##STR2421##
##STR2422## ##STR2423## CH.sub.3 ##STR2424## ##STR2425##
##STR2426## CH.sub.3 ##STR2427## ##STR2428## ##STR2429## CH.sub.3
##STR2430## ##STR2431## ##STR2432## CH.sub.3 ##STR2433##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula VII' ##STR2434## where the substituent
R.sup.4 and R.sup.4' are hydrogen or lower alkyl, and R.sup.24,
R.sup.25 and W are selected according to Table VII.
TABLE VII' ______________________________________ W R.sup.24
R.sup.25 ______________________________________ ##STR2435##
CH.sub.3 ##STR2436## ##STR2437## CH.sub.3 ##STR2438## ##STR2439##
CH.sub.3 ##STR2440## ##STR2441## CH.sub.3 ##STR2442## ##STR2443##
CH.sub.3 ##STR2444## ##STR2445## CH.sub.3 ##STR2446## ##STR2447##
CH.sub.3 ##STR2448## ##STR2449## CH.sub.3 ##STR2450## ##STR2451##
CH.sub.3 ##STR2452## ##STR2453## CH.sub.3 ##STR2454## ##STR2455##
CH.sub.3 ##STR2456## ##STR2457## CH.sub.3 ##STR2458## ##STR2459##
CH.sub.3 ##STR2460## ##STR2461## CH.sub.3 ##STR2462## ##STR2463##
CH.sub.3 ##STR2464## ##STR2465## CH.sub.3 ##STR2466## ##STR2467##
CH.sub.3 ##STR2468## ##STR2469## CH.sub.3 ##STR2470## ##STR2471##
CH.sub.3 ##STR2472## ##STR2473## CH.sub.3 ##STR2474## ##STR2475##
CH.sub.3 ##STR2476## ##STR2477## CH.sub.3 ##STR2478## ##STR2479##
CH.sub.3 ##STR2480## ##STR2481## CH.sub.3 ##STR2482## ##STR2483##
CH.sub.3 ##STR2484## ##STR2485## CH.sub.3 ##STR2486## ##STR2487##
CH.sub.3 ##STR2488## ##STR2489## CH.sub.3 ##STR2490## ##STR2491##
CH.sub.3 ##STR2492## ##STR2493## CH.sub.3 ##STR2494## ##STR2495##
CH.sub.3 ##STR2496## ##STR2497## CH.sub.3 ##STR2498## ##STR2499##
CH.sub.3 ##STR2500## ##STR2501## CH.sub.3 ##STR2502## ##STR2503##
CH.sub.3 ##STR2504## ##STR2505## CH.sub.3 ##STR2506## ##STR2507##
CH.sub.3 ##STR2508## ##STR2509## CH.sub.3 ##STR2510## ##STR2511##
CH.sub.3 ##STR2512## ##STR2513## CH.sub.3 ##STR2514## ##STR2515##
CH.sub.3 ##STR2516## ##STR2517## CH.sub.3 ##STR2518## ##STR2519##
CH.sub.3 ##STR2520## ##STR2521## CH.sub.3 ##STR2522## ##STR2523##
CH.sub.3 ##STR2524## ##STR2525## CH.sub.3 ##STR2526## ##STR2527##
CH.sub.3 ##STR2528## ##STR2529## CH.sub.3 ##STR2530## ##STR2531##
CH.sub.3 ##STR2532## ##STR2533## CH.sub.3 ##STR2534## ##STR2535##
CH.sub.3 ##STR2536## ##STR2537## CH.sub.3 ##STR2538## ##STR2539##
CH.sub.3 ##STR2540## ##STR2541## CH.sub.3 ##STR2542## ##STR2543##
CH.sub.3 ##STR2544## ##STR2545## CH.sub.3 ##STR2546## ##STR2547##
CH.sub.3 ##STR2548## ##STR2549## CH.sub.3 ##STR2550## ##STR2551##
CH.sub.3 ##STR2552## ##STR2553## CH.sub.3 ##STR2554## ##STR2555##
CH.sub.3 ##STR2556## ##STR2557## CH.sub.3 ##STR2558## ##STR2559##
CH.sub.3 ##STR2560## ##STR2561## CH.sub.3 ##STR2562## ##STR2563##
CH.sub.3 ##STR2564## ##STR2565## CH.sub.3 ##STR2566## ##STR2567##
CH.sub.3 ##STR2568## ##STR2569## CH.sub.3 ##STR2570## ##STR2571##
CH.sub.3 ##STR2572## ##STR2573## CH.sub.3 ##STR2574## ##STR2575##
CH.sub.3 ##STR2576## ##STR2577## CH.sub.3 ##STR2578## ##STR2579##
CH.sub.3 ##STR2580## ##STR2581## CH.sub.3 ##STR2582## ##STR2583##
CH.sub.3 ##STR2584## ##STR2585## CH.sub.3 ##STR2586## ##STR2587##
CH.sub.3 ##STR2588## ##STR2589## CH.sub.3 ##STR2590## ##STR2591##
CH.sub.3 ##STR2592## ##STR2593## CH.sub.3 ##STR2594## ##STR2595##
CH.sub.3 ##STR2596## ##STR2597## CH.sub.3 ##STR2598## ##STR2599##
CH.sub.3 ##STR2600## ##STR2601## CH.sub.3 ##STR2602## ##STR2603##
CH.sub.3 ##STR2604## ##STR2605## CH.sub.3 ##STR2606## ##STR2607##
CH.sub.3 ##STR2608## ##STR2609## CH.sub.3 ##STR2610## An
alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula VIII' ##STR2611## VIII' where the
substituents R and R' are hydrogen, halo or perfluro-loweralkyl,
R.sup.4 is hydrogen or lower alkyl, and R.sup.24, R.sup.25, V, and
##STR2612## are selected according to Table VIII'.
TABLE VIII'
__________________________________________________________________________
##STR2613## ##STR2614## ##STR2615## ##STR2616##
__________________________________________________________________________
COOH ##STR2617## CH.sub.3 ##STR2618## COOH ##STR2619## CH.sub.3
##STR2620## COOCH.sub.3 ##STR2621## CH.sub.3 ##STR2622##
COOCH.sub.3 ##STR2623## CH.sub.3 ##STR2624## COOC.sub.2 H.sub.5
##STR2625## CH.sub.3 ##STR2626## ##STR2627## ##STR2628## CH.sub.3
##STR2629## ##STR2630## ##STR2631## CH.sub.3 ##STR2632## COOH
##STR2633## CH.sub.3 ##STR2634## ##STR2635## ##STR2636## CH.sub.3
##STR2637## COOCH.sub.3 ##STR2638## CH.sub.3 ##STR2639## COOH
##STR2640## CH.sub.3 ##STR2641## COOH ##STR2642## CH.sub.3
##STR2643## COOH ##STR2644## CH.sub.3 ##STR2645## ##STR2646##
##STR2647## CH.sub.3 ##STR2648## COOCH.sub.3 ##STR2649## CH.sub.3
##STR2650## ##STR2651## ##STR2652## CH.sub.3 ##STR2653## COOH
##STR2654## CH.sub.3 ##STR2655## COOH ##STR2656## CH.sub.3
##STR2657## COOCH.sub.3 ##STR2658## CH.sub.3 ##STR2659##
COOCH.sub.3 ##STR2660## CH.sub.3 ##STR2661## COOCH.sub.2 H.sub.5
##STR2662## CH.sub.3 ##STR2663## ##STR2664## ##STR2665## CH.sub.3
##STR2666## ##STR2667## ##STR2668## CH.sub.3 ##STR2669## COOH
##STR2670## CH.sub.3 ##STR2671## ##STR2672## ##STR2673## CH.sub.3
##STR2674## COOCH.sub.3 ##STR2675## CH.sub.3 ##STR2676## COOH
##STR2677## CH.sub.3 ##STR2678## COOH ##STR2679## CH.sub.3
##STR2680## COOH ##STR2681## CH.sub.3 ##STR2682## ##STR2683##
##STR2684## CH.sub.3 ##STR2685## COOCH.sub.3 ##STR2686## CH.sub.3
##STR2687## ##STR2688## ##STR2689## CH.sub.3 ##STR2690## COOH
##STR2691## CH.sub.3 ##STR2692## COOH ##STR2693## CH.sub.3
##STR2694## COOCH.sub.3 ##STR2695## CH.sub.3 ##STR2696##
COOCH.sub.3 ##STR2697## CH.sub.3 ##STR2698## COOC.sub.2 H.sub.5
##STR2699## CH.sub.3 ##STR2700## ##STR2701## ##STR2702## CH.sub.3
##STR2703## ##STR2704## ##STR2705## CH.sub.3 ##STR2706## COOH
##STR2707## CH.sub.3 ##STR2708## ##STR2709## ##STR2710## CH.sub.3
##STR2711## COOCH.sub.3 ##STR2712## CH.sub.3 ##STR2713## COOH
##STR2714## CH.sub.3 ##STR2715## COOH ##STR2716## CH.sub.3
##STR2717## COOH ##STR2718## CH.sub.3 ##STR2719## ##STR2720##
##STR2721## CH.sub.3 ##STR2722## COOCH.sub.3 ##STR2723## CH.sub.3
##STR2724## ##STR2725## ##STR2726## CH.sub.3 ##STR2727## COOH
##STR2728## C.sub.2 H.sub.5 ##STR2729## COOH ##STR2730## C.sub.2
H.sub.5 ##STR2731## COOCH.sub.3 ##STR2732## C.sub.2 H.sub.5
##STR2733## COOCH.sub.3 ##STR2734## C.sub.2 H.sub.5 ##STR2735##
COOC.sub.2 H.sub.5 ##STR2736## C.sub.2 H.sub.5 ##STR2737##
##STR2738## ##STR2739## C.sub.2 H.sub.5 ##STR2740## ##STR2741##
##STR2742## C.sub.2 H.sub.5 ##STR2743## COOH ##STR2744## C.sub.2
H.sub.5 ##STR2745## ##STR2746## ##STR2747## C.sub.2 H.sub.5
##STR2748## COOCH.sub.3 ##STR2749## C.sub.2 H.sub.5 ##STR2750##
COOH ##STR2751## C.sub.2 H.sub.5 ##STR2752## COOH ##STR2753##
C.sub.2 H.sub.5 ##STR2754## COOH ##STR2755## C.sub.2 H.sub.5
##STR2756##
##STR2757## ##STR2758## C.sub.2 H.sub. 5 ##STR2759## COOCH.sub.3
##STR2760## C.sub.2 H.sub.5 ##STR2761## ##STR2762## ##STR2763##
C.sub.2 H.sub.5 ##STR2764## COOH ##STR2765## CH.sub.3 ##STR2766##
COOH ##STR2767## CH.sub.3 ##STR2768## COOCH.sub.3 ##STR2769##
CH.sub.3 ##STR2770## COOCH.sub.3 ##STR2771## CH.sub.3 ##STR2772##
COOC.sub.2 H.sub.5 ##STR2773## CH.sub.3 ##STR2774## ##STR2775##
##STR2776## CH.sub.3 ##STR2777## ##STR2778## ##STR2779## CH.sub.3
##STR2780## COOH ##STR2781## CH.sub.3 ##STR2782## ##STR2783##
##STR2784## CH.sub.3 ##STR2785## COOCH.sub.3 ##STR2786## CH.sub.3
##STR2787## COOH ##STR2788## CH.sub.3 ##STR2789## COOH ##STR2790##
CH.sub.3 ##STR2791## COOH ##STR2792## CH.sub.3 ##STR2793##
##STR2794## ##STR2795## CH.sub.3 ##STR2796## COOCH.sub.3
##STR2797## CH.sub.3 ##STR2798## ##STR2799## ##STR2800## CH.sub.3
##STR2801## COOH ##STR2802## ##STR2803## ##STR2804## COOH
##STR2805## ##STR2806## ##STR2807## COOCH.sub.3 ##STR2808##
##STR2809## ##STR2810## COOCH.sub.3 ##STR2811## ##STR2812##
##STR2813## COOC.sub.2 H.sub.5 ##STR2814## ##STR2815## ##STR2816##
##STR2817## ##STR2818## ##STR2819## ##STR2820## ##STR2821##
##STR2822## ##STR2823## ##STR2824## COOH ##STR2825## ##STR2826##
##STR2827## ##STR2828## ##STR2829## ##STR2830## ##STR2831##
COOCH.sub.3 ##STR2832## ##STR2833## ##STR2834## COOH ##STR2835##
##STR2836## ##STR2837## COOH ##STR2838## ##STR2839## ##STR2840##
COOH ##STR2841## ##STR2842## ##STR2843## ##STR2844## ##STR2845##
##STR2846## ##STR2847## COOCH.sub.3 ##STR2848## ##STR2849##
##STR2850## ##STR2851## ##STR2852## ##STR2853## ##STR2854## COOH
##STR2855## CH.sub.3 ##STR2856## COOH ##STR2857## CH.sub.3
##STR2858## COOCH.sub.3 ##STR2859## CH.sub.3 ##STR2860##
COOCH.sub.3 ##STR2861## CH.sub.3 ##STR2862## COOC.sub.2 H.sub.5
##STR2863## CH.sub.3 ##STR2864## ##STR2865## ##STR2866## CH.sub.3
##STR2867## ##STR2868## ##STR2869## CH.sub.3 ##STR2870## COOH
##STR2871## CH.sub.3 ##STR2872## ##STR2873## ##STR2874## CH.sub.3
##STR2875## COOCH.sub.3 ##STR2876## CH.sub.3 ##STR2877## COOH
##STR2878## CH.sub.3 ##STR2879## COOH ##STR2880## CH.sub.3
##STR2881## COOH ##STR2882## CH.sub.3 ##STR2883## ##STR2884##
##STR2885## CH.sub.3 ##STR2886## COOCH.sub.3 ##STR2887## CH.sub.3
##STR2888## ##STR2889## ##STR2890## CH.sub.3 ##STR2891## COOH
##STR2892## CH.sub.3 ##STR2893## COOH ##STR2894## CH.sub.3
##STR2895## COOCH.sub.3 ##STR2896## CH.sub.3 ##STR2897##
COOCH.sub.3 ##STR2898## CH.sub.3 ##STR2899## COOC.sub.2 H.sub.5
##STR2900## CH.sub.3 ##STR2901## ##STR2902## ##STR2903## CH.sub.3
##STR2904## ##STR2905## ##STR2906## CH.sub.3 ##STR2907## COOH
##STR2908## CH.sub.3 ##STR2909## ##STR2910## ##STR2911## CH.sub.3
##STR2912## COOCH.sub.3 ##STR2913## CH.sub.3 ##STR2914## COOH
##STR2915## CH.sub.3 ##STR2916## COOH ##STR2917## CH.sub.3
##STR2918## COOH ##STR2919## CH.sub.3 ##STR2920## ##STR2921##
##STR2922## CH.sub.3 ##STR2923## COOCH.sub.3 ##STR2924## CH.sub.3
##STR2925## ##STR2926## ##STR2927## CH.sub.3 ##STR2928## COOH
##STR2929## C.sub.4 H.sub.9 ##STR2930## COOH ##STR2931## C.sub.4
H.sub.9 ##STR2932## COOCH.sub.3 ##STR2933## C.sub.4 H.sub.9
##STR2934## COOCH.sub.3 ##STR2935## C.sub.4 H.sub.9 ##STR2936##
COOC.sub.2 H.sub.5 ##STR2937## C.sub.4 H.sub.9 ##STR2938##
##STR2939## ##STR2940## C.sub.4 H.sub.9 ##STR2941## ##STR2942##
##STR2943## C.sub.4 H.sub.9 ##STR2944## COOH ##STR2945## C.sub.4
H.sub.9 ##STR2946## ##STR2947## ##STR2948## C.sub.4 H.sub.9
##STR2949## COOCH.sub.3 ##STR2950## C.sub.4 H.sub.9 ##STR2951##
COOH ##STR2952## C.sub.4 H.sub.9 ##STR2953## COOH ##STR2954##
C.sub.4 H.sub.9 ##STR2955## COOH ##STR2956## C.sub.4 H.sub.9
##STR2957## ##STR2958## ##STR2959## C.sub.4 H.sub.9 ##STR2960##
COOCH.sub.3 ##STR2961## C.sub.4 H.sub.9 ##STR2962## ##STR2963##
##STR2964## C.sub.4 H.sub.9 ##STR2965##
__________________________________________________________________________
An alternative preferred embodiment of the invention comprises a
compound capable of inhibiting farnesyl:protein transferase at a
concentration equal to or lower than that of the tetrapeptide CVFM
represented by Formula IX ##STR2966## where the substituents R and
R' are halo or perfluro-loweralkyl, R.sup.1 is CF.sub.3 or phenyl
optionally substituted with halo or haloloweralkyl, R.sup.4,
R.sup.4', and R.sup.28 are hydrogen or lower alkyl, W is
--(C.dbd.O)--NR.sup.7' R.sup.8 where R.sup.7' and R.sup.8 are
selected according to Table Ia', R.sup.25' is C.sub.1 -C.sub.4
alkyl-SH or C.sub.1 -C.sub.4 alkyl-S-C.sub.1 -C.sub.4 alkyl, and
##STR2967## represents methylene, ethylene, or ethenylene,
optionally substituted with oxo (.dbd.O), loweralkyl, or
haloloweralkyl. Subgeneric preferred examples of IX include;
##STR2968## where the substituents are as described above.
D. Methods of Making
In the schemes and examples below, the following standard
abbreviations are employed.
______________________________________ ABBREVIATIONS
______________________________________ Boc or BOC
tert-butyloxycarbonyl Fmoc or fluorenylmethyloxycarbonyl FMOC DCC
dicyclohexylcarbodiimide PyBrOP
bromo-tri-pyrrolidinophosphoniumhexafluoro- phosphate TFA
trifluoroacetic acid DIPC diisopropylcarbodiimide BOP
benzotriazolyloxy-trisdimethylamino-phosphonium hexafluorophosphate
EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
DEAD diethylazodicarboxylate Ph.sub.3 P triphenylphosphine LDA
lithium diisopropylamine NMM N-methylmorpholine EtN(iPr).sub.2
diisopropylethylamine SPPS solid phase peptide synthesis
t-BuO.sub.2 C tert-butoxycarbonyl DMF dimethylformamide DTT
dithiothreitol DMA dimethylacetamide DMAP dimethylaminopyridine DCM
dichloromethane DMSO dimethylsulfoxide DIPEA diisopropylethylamine
DMEM Dulbecco's modified essential media MBHA methylbenzhydryamine
ACN acetonitrile HPLC high pressure liquid chromatography
______________________________________
General methods of synthesis of the compounds of this invention are
shown in Schemes I-XIV. Compounds bearing a 3-amino substituent in
the benzodiazepine ring are synthesized as shown in Schemes I-III.
Typically, a triply convergent route is employed, which joins the
key intermediates 9 or 10 (Scheme I) with suitably functionalized
amine and carboxyl components (Schemes II and III) using standard
amide bond-forming procedures.
As shown in Scheme I, the protected amino acids 9 and 10 may be
prepared from a suitably substituted 2-aminobenzophenone (1). Many
2-aminobenzophenones are known in the art or are available from
commercial sources such as Aldrich Chemical Co. General methods for
the synthesis of new 2-aminobenzophenones may be found in the
literature (c.f. Walsh, D. A. Synthesis, 1980, 677-688).
##STR2969##
Acylation of 1 with a haloacetyl halide, such as bromoacetyl
bromide in a suitable solvent mixture, such as water/CH.sub.2
Cl.sub.2, typically at temperatures ranging from 0.degree. C. to
25.degree. C., produces amide 2. Reaction of 2 with ammonia in a
polar solvent such as methanol at 25.degree. to 75.degree. C. then
gives the 1,4-benzodiazepin-2-one 3, after evaporation of the
solvent. Alkylation of 3 with a substituted organic ester (4),
preferably tert-butyl bromoacetate, in the presence of a base,
preferably Cs.sub.2 CO.sub.3 in 1-methyl-2-pyrrolidinone at ambient
temperature, gives 5. Alternatively, 3 may be alkylated at N-1 with
a variety of other alkylating agents, for instance, esters of
substituted or unsubstituted acrylates, 4-bromobutanoates, etc.
Branched compounds (i.e. R.sup.4 and/or R.sup.4' .noteq.H), may be
prepared by generation of the polyanion of 5 with base and
alkylation with an appropriate alkyl halide.
Subsequent to alkylation, the ester of 5 may be cleaved with an
acid such as TFA (for the tert-butyl esters) or under mild aqueous
base hydrolysis (for other alkyl esters) at temperatures between
0.degree. and 25.degree. C.
The acid 6 is converted to amino acid 8 via reaction of the
dianion, generated with at least two equivalents of a strong base
with an electrophilic aminating agent. Alternatively, 6 may be
halogenated and reacted with an amine source such as azide
(followed by reduction) or ammonia. Preferably, 6 is reacted with 4
equivalents of potassium tert-butoxide in glyme at -5.degree. C.
for 30 min and treated with 1.1 equivalents of isobutyl nitrite.
The resulting oxime 7 can then be reduced to the racemic amino acid
8 using a variety of reductants, preferably hydrogenation at 40
psig in the presence of Ruthenium on carbon or Raney nickel in
methanol at 50.degree. to 70.degree. C. for 1-4 days.
Amino acid 8 is then suitably protected for selective coupling at
the carboxyl terminus. For example, 8 can be converted to the N-BOC
derivitive 9 using standard amino acid protection conditions,
preferably, reaction with equimolar amounts of di-tert-butyl
dicarbonate and triethyl amine in DMF/water at ambient
temperature.
For compounds where R.sup.24 .noteq.H, 9 can be alkylated at
nitrogen with a wide variety of alkylating agents including
n-alkyl, branched alkyl, and benzyl, according to the standard
procedure of Benoiton, et al., Can. J. Chem. 1977, 55, 906. For
example, reaction of 9 with at least 2 equivalents of base and an
alkylating agent in a polar, aprotic solvent at 0.degree. to
50.degree. C. for 0.5 to 48 h gives 10. Preferably, reaction with 3
equivalents of sodium hydride and 4 equivalents of methyl iodide in
THF at -5.degree. to 5.degree. C. gives 10 (R.sup.24 =Me).
Compounds 9 and 10 can be further elaborated according to Schemes
II and III. In general, the carboxylic acid function of 9 and 10 is
reacted with a suitably protected amine component using standard
solid phase (Scheme II) or solution phase (Scheme III) peptide
synthesis procedures. The BOC or other protecting group on N-3 of
the benzodiazapinone is removed and the amine function then coupled
with a third component, for example, a suitably protected amino
acid, and then deprotected, again employing standard procedures.
The resulting product is subsequently purified by chromatography or
crystallization. ##STR2970##
Once fully deprotected, compounds of the type 17 and 23, and salts
thereof, may be further modified at the carboxy and/or amino
terminus by esterification or acylation, respectively, employing
standard procedures. ##STR2971##
The synthesis of a second class of compounds of this invention is
shown in Scheme IV. Acylation of the 2-amino-benzophenones 1 with
an N-protected (preferably BOC) amino acid using DCC or the mixed
anhydride method gives 25. A wide variety of protected amino acids
24 may be used in this reaction, including side chain protected
natural amino acids (both D and L), substituted phenyl glycines,
thiolysine, and the variety of synthetic, non-natural amino acids
known to one skilled in the art (see e.g. U.S. Pat. No. 5,120,859,
WO 93/04081 and 37 CFR 1.822(b)(2) and 1.822(p)(2)). Preferably,
the side chain functions of the amino acid are protected
orthogonally to the alpha-amine to facilitate selective
deprotection. Treatment of the deblocked compound with base,
preferably in methanol, gives the 3-substituted benzodiazepin-2-one
26. Alkylation at N-1 with a substituted ester as described for 3
(Scheme I) gives 7 which is deprotected and coupled to a protected
amino acid using standard procedures to afford 29. Final deblocking
of the side chain protecting groups and purification gives 30.
##STR2972##
Alternatively, 26 may be directly alkylated with the "top"
sidechain in one intact piece, as shown in Scheme V. Reaction of 26
with an alkyl halide such as a suitably substituted benzyl bromide,
alkyl bromide, in the presence of a base, preferably Nail or
Cs.sub.2 CO.sub.3, gives 31. Deprotection under standard conditions
and purification affords 32. ##STR2973##
When the amino acid side chain of 32 is that of serine, futher
manipulation is possible as described in Scheme VI. Deprotection of
the hydroxyl function gives free alcohol 34 which can be alkylated
or acylated at oxygen to give 35 using standard ether synthesis or
acylation procedures. Compounds of the type 32, with the side chain
of cysteine are treated in an analogous fashion to 33.
Alternatively, 34 may be converted to amine 36 under Mitsunobu
conditions, preferably using Ph.sub.3 P, diethyl azodicarboxylate
(DEAD), and HN.sub.3. Reduction of the resulting azide, preferably
by hydrogenation over Pd/C, gives amine 36. 36 is then alkylated or
acylated to give 37 and 38, respectively, after deprotection and
purification. ##STR2974##
For compounds 32, with the side chains derived from aspartic or
glutamic acids, further modification is carried out as per Scheme
VII. Thus, selective removal of the side chain ester function,
preferably benzyl or lower alkyl, using appropriate conditions,
preferably aqueous NaOH, gives the free acid. Coupling to a second
amine component, such as 2-mercaptoethyl amine, using standard
conditions, preferably DCC, gives the protected amide which is
fully deprotected and purified to afford 40. ##STR2975##
Another variety of compounds which are the subject of this
invention are synthesized as shown in Scheme VIII. Alkylation at
N-1 of benzodiazepinone 3 (as described for 31, Scheme V) gives 41.
This alkylation may, for example, be conducted with any halo
substituted loweralkyl, loweralkylaryl, or loweralkylheterocycle
(--CR.sup.4 R.sup.4' --W-- in Scheme VIII). The alkyl, aryl, or
heterocycle moieties may be substituted with protected (-P.G.)
carboxyls, tetrazoles, thiols, etc, or precursors of these groups
(e.g. nitriles for tetrazoles, etc.). Deprotonation at C-3 of the
heterocycle, preferably with LDA in THF at less than 50.degree. C.,
gives anion 42 which can be reacted with a variety of
electrophiles. For example, reaction with substituted aldehydes,
active esters, and alkyl halides gives, after deprotection and
purification, products 43, 44, and 45, respectively.
##STR2976##
In cases where the starting materials are optically active, the
chirality at C-3 of the benzodiazepinone is controlled by the
starting materials. When racemic starting materials are employed,
diastereomeric products are obtained. The diastereomers may be
separated by chromatography.
Benzodiazepines of the instant invention with a spiro linkage at
C-3 may be made according to Scheme IX. The C-3 amine is first
coupled to a 9-phenylfluorenyl protected amino acid to give 48,
followed by reaction with a dihalo substituted alkane in base to
give 49. The 9-phenylfluorenyl group is then replaced with BOC and
the resulting compound 50 is reacted with an immobilized free amine
(e.g. compound 12, Scheme II) in a solid phase synthesis procedure.
##STR2977##
Benzodiazepines of the instant invention with a hydroxamic or
carboxylic terminus at C-3 may be made according to Scheme X.
Compound 14 from Scheme II may be treated with an acid anhydride in
N-methyl-morpholine (NMM) to produce 52 contemplated to be a
suitable ras FT inhibitor. 52 in turn may be coupled to a
hydroxamic acid and cleaved to produce 54. ##STR2978##
Benzodiazepine compounds 16 and 22 of Schemes II and III, where
R.sup.24 is hydrogen may be convereted to heterocycles of formulae
IXa-IXd according to Scheme XI. Here, R.sup.25' and W are suitably
blocked prior to reacting with the cyclizing agent, followed by
deblocking and release from the resin as described in Schemes II
and III. ##STR2979##
The imine nitrogen of the instant benzodiazepines may be cyclized
with an amide nitrogen bonded to C-3 by first reacting 63 with
bromoacetyl bromide followed by reduction of the imine and closure
to give 64. ##STR2980##
The synthesis of a class of compounds represented by formula (V) is
shown in Scheme XIII. Reductive alkylation of the
2-amino-benzophenones 1 with an N-protected (preferably BOC)
aldehyde using NaCNBH.sub.3 gives 65. A wide variety of aldehydes
derived from protected amino acids may be used in this reaction,
including side chain protected natural amino acids (both D and L),
substituted phenyl glycines, thiolysine, and the variety of
synthetic, non-natural amino acids known to one skilled in the art.
65 is then acylated with an N-protected (preferably Fmoc) amino
acid in the presence of DCC (see Scheme IV) to afford 66. 66 is
then treated with base to form the benzodiazepine 67. This
benzodiazepine is treated with Lawessons Reagent (Fluka) followed
by MeI and then anhydrous HCl, followed by neutralization and
heating to give the tricyclic compound 68. The optionally protected
sidechain is deprotected to give 69. ##STR2981##
Compounds bearing a 3-ureido, 3-carbamoyl, or 3-thiocarbamoyl
substituent on the benzodiazepine ring are synthesized as shown in
Scheme XIV. Compound 9 from Scheme I may be esterified and reacted
with TFA to yield 70. The 3-amino group is then converted to the
isocyanate 71 with phosgene or 1,1'-carbonyldiimidazole. 71 may be
converted into the corresponding ureido, carbamoyl, or
thiocarbamoyl, by reacting it with suitably protected amines,
alcohols, or thioIs yielding 72, 73, and 74 respectively. A wide
variety of protected amino acids having a free amino group are
prefered in this reaction, including side chain protected natural
amino acids (both D and L), substituted phenyl glycines,
thiolysine, and a variety of synthetic, non-natural amino acids
(e.g. thioproline, .beta.-alanine, etc.) known to one skilled in
the art (see e.g. U.S. Pat. No. 5,120,859, and WO 93/04081). Simple
suitably protected alkylaminothiols and hydroxyalkylthiols may
similarly be employed to produce 72, 73, or 74. ##STR2982##
72, 73, or 74 may be used as inhibitors per se or further converted
into compounds 75, 76, or 77 by coupling protected amino acids to
the free acid forms of 72, 73, or 74 using the standard procedures
shown above.
EXAMPLES
In Examples 1-8 the parenthetical compound numbers refer to the
numbers in Scheme 1 below. ##STR2983##
EXAMPLE 1
2,3,4,5-Tetrahydro-1H-[1]-benzazapin-2-one (1)
To a stirred suspension of 13.4 g (207 mmol) of sodium azide and
25.0 ml (188 mmol) of .alpha.-tetralone in 150 mL of chloroform,
was added 50.0 mL of concentrated sulfuric acid, dropwise over 1 h.
After 30 min., the chloroform phase was decanted and the acidic
phase poured into 1 L of water. The precipitated solid was
collected on a filter, washed with water, and recrystallized from 1
L of boiling water. The product was collected and dried under
vacuum to yield 14.3 g (47%) of tan needles. .sup.1 H NMR (300 MHz,
CDCl.sub.3) d 8.63 (1H, bs), 7.24 (2H, m), 7.13 (1H, m), 7.03 (1H,
d, J=8 Hz), 2.82 (2H, t, J=6 Hz), 2.38 (2H, t, J=6 Hz), 2.24 (2H,
m).
EXAMPLE 2
3-Iodo-2,3,4,5 -tetrahydro-1H-[1]-benzazapin-2-one (2)
A suspension of 34.2 g (212 mmol) of
2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one and 224 mL (171.2 g,
1.06 mol) of hexamethyldisilazane in 400 mL of methylene chloride
was heated at reflux for 15 min and cooled to 30.degree. C. Iodine
(161.5 g, 636 mmol) was added in one portion, the solution heated
at reflux for 2.5 h, cooled, and poured into a 0.degree. C.
solution of 88.6 g of sodium sulfite in 800 mL of water, with
vigorous stirring. The aqueous phase was separated, extracted with
methylene choride and the combined organics were washed with water
and concentrated in vacuo to approximately 200 mL. Toluene (800 mL)
was added, the solution was concentrated to a slurry, and the
product collected on a filter. Drying under vacuum gave 36.7 g
(60%) of a tan powder. .sup.1 H NMR (300 MHz, CDCl.sub.3) d 8.47
(1H, bs), 7.3-7.1 (3H, m), 7.06 (1H, d, J=8 Hz), 4.68 (1H, t, J=8.7
Hz), 2.97 (1H, m), 2.80-2.60 (3H, m).
EXAMPLE 3
3-Azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one (3)
To a solution of 36.7 g (128 mmol) of
3-Iodo-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one in 200 mL of
dimethylformamide was added 9.97 g (153 mmol) of sodium azide.
After 3 h, the mixture was poured into 800 mL of ice water and the
precipitate collected on a filter. After washing the solid
successively with water, 3% aqueous sodium bisulfite, and water,
the product was dried under vacuum to give 21.5 g (83%) of a tan
powder. .sup.1 H NMR (300 MHz, CDCl.sub.3) d 8.91 (1H, bs), 7.4-7.0
(4H, m), 3.89 (1H, t, J=9 Hz), 2.97 (1H, m), 2.73 (1H, m), 2.52
(1H, m), 2.32 (1H, m).
EXAMPLE 4
Ethyl 3-azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-acetate
(4)
To a solution of 5.00 g (24.7 mmol) of
3-Azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one, 1.48 g (26.5
mmol) of powdered potassium hydroxide, and 780 mg (2.47 mmol) of
tetrabutylammonium bromide in 25 mL of tetrahydrofuran was added
2.95 mL (4.42 g, 26.5 mmol) of ethyl bromoacetate. The mixture was
rapidly stirred at ambient temperature for 4 h and partitioned
between ethyl acetate and water. The organic phase was washed with
water, dried (magnesium sulfate), and concentrated, to yield 6.43 g
of an oil, used without further purification. .sup.1 H NMR (300
MHz, CDCl.sub.3) d 7.38-7.13 (4H, m), 4.72 (1H, d, J=17 Hz), 4.42
(1H, d, J=17 Hz), 4.18 (2H, q, J=7 Hz), 3.75 (1H, m), 3.38 (1H, m),
2.70 (1H, m), 2.40 (2H, m), 1.26 (3H, t, J=7 Hz).
EXAMPLE 5
Ethyl
3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzaza-pin-2-one-1
-acetate (5)
A suspension of 6.43 g of crude ethyl
3-azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-acetate and 1
g of 10% palladium on carbon in 40 mL of ethanol was shaken under
50 psig H.sub.2 for 12 h. The mixture was filtered through celite,
concentrated to a foam, and redissolved in 100 ml of methylene
chloride/1N aqueous sodium bicarbonate (1:1). Di-t-butyldicarbonate
(10.8 g, 49.4 mmol) was added, the mixture was rapidly stirred at
ambient temperature for 12 h, and partitioned between water and
methylene chloride. The organic phase was separated and washed
successively with 1N sodium bicarbonate, 1N sodium bisulfate,
water, brine, and dried over magnesium sulfate. Concentration in
vacuo gave a solid that was chromatographed (200 g silica gel 60,
ethyl acetate/hexane 1:3 to 1:2). Recrystallization from ethyl
acetate/hexane gave 5.93 g (69% from
3-azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one) of a colorless
crystalline solid. .sup.1 H NMR (300 MHz, CDCl.sub.3) d 7.3-7.1
(4H, m), 5.42 (1H, bd), 4.75 (1H, d, J=17 Hz), 4.33 (1H, d, J=17
Hz), 4.25 (1H, m), 4.17 (1H, bq, J=7 Hz), 3.32 (1H, m), 2.57 (2H,
m), 1.98 (1H, m), 1.38 (9H, s), 1.24 (3H, t, J=7 Hz). Exact mass
(FAB, M+H.sup.+) calcd for C.sub.19 H.sub.27 N.sub.2 O.sub.5 :
363.1920; Found: 363.1929.
EXAMPLE 6
3-(tert-Butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-a
cetic acid (6)
A solution of 5.00 g (14.3 mmol) of ethyl
3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-
acetate in 30 mL of methanol was cooled to 0.degree. C. and treated
with 28.5 mL of 1N sodium hydroxide. Tetrahydrofuran was added
until the mixture was homogeneous (about 10 mL) and the solution
warmed to ambient temperature for 3 h. The volatiles were removed
in vacuo and the residue was diluted with water and extracted with
ether. The aqueous phase was acidified to pH 2 with 1N sodium
bisulfate and extracted twice with ethyl acetate. The combined
organics were washed with brine, dried over magnesium sulfate, and
concentrated to 20 mL. Hexane was added and the resulting
suspension was aged overnight at 0.degree. C., filtered, and the
solid dried in vacuo to give 4.62 g (100%) of colorless product.
.sup.1 H NMR (300 MHz, CDCl.sub.3) d 8.65 (1H, b), 7.3-7.1 (4 H,
m), 5.52 (1H, bd), 4.71 (1H, d, J=17 Hz), 4.40 (1H, d, J=17 Hz),
4.25 (1H, m), 3.25 (1H, m), 2.56 (2H, m), 1.98 (1H, m), 1.37 (9H,
s). Exact mass (FAB, M+H.sup.+) calcd for C.sub.17 H.sub.23 N.sub.2
O.sub.5 : 335.1607; Found: 335.1609.
EXAMPLE 7
Methyl
3-(tert-butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-
one-1-acetate (7)
To a solution of 2.58 g (8.00 mmol) of
3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-
acetic acid in 40 mL of tetrahydrofuran/dimethylformamide/glyme
(6:1:1) was added 3.98 mL (64.0 mmol) of methyl iodide and 960 mg
of sodium hydride (60% dispersion in mineral oil, 24.0 mmol). The
suspension was heated at 50.degree. C. (at which point it became
homogeneous) for 3 h and cooled. 30 mL of 1N sodium bisulfate was
added, the volatiles removed in vacuo, and the aqueous slurry
extracted twice with ethyl acetate. The organic phase was washed
with brine, dried over magnesium sulfate, filtered, and
concentrated. Chromatography (150 g silica gel 60, ethyl
acetate/hexane/acetic acid (40:60:1)) gave a solid that was
recrystallized from methanol/water to yield 2.01 g (69%) of
colorless product. .sup.1 H NMR (300 MHz, CDCl.sub.3, spectrum
broad due to carbamate rotomers) d 7.3-7.1 (4H, m), 4.95 -4.15 (3H,
bm), 3.69 (3H, bs), 3.35 (1H, m), 3.02 (3H, s), 2.68 (1H, m), 2.50
(1H, m), 2.15 (1H, m), 1.39 (4.5H, bs), 1.29 (4.5H, bs). Mass spec.
(FAB, M+H.sup.+) calcd for C.sub.19 H.sub.27 N.sub.2 O.sub.5 :
363.19; Found: 363.1.
EXAMPLE 8
3-(tert-Butoxycarbonylmethylamino
)-2,3,4,5-tetrahydro-1H-[1]-benz-azapin-2-one-1-acetic acid (8)
To a 0.degree. C. methanolic solution of 1.59 g (4.39 mmol) of
methyl
3-(tert-butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-
one-1-acetate was added 8.6 mL of 1N sodium hydroxide and 5 mL of
tetrahydrofuran. The mixture was magnetically stirred for 3 h at
ambient temperature and concentrated in vacuo to remove the
volatiles. The slurry was diluted with water, extracted with ether
(discarded), and acidified to pH 2 with 1N sodium hydrogen sulfate.
The aqueous phase was extracted twice with ethyl acetate and the
combined organics were washed with brine, dried over magnesium
sulfate, filtered, and concentrated to a colorless foam. .sup.1 H
NMR (300 MHz, CDCl.sub.3) d 8.40 (1H, b), 7.35-7.05 (4H, m), 4.80
(1H, m), 4.50 (1H, m), 4.25 (1H, m), 3.25 (1H, m), 3.00 (3H, s),
2.65 (1H, m), 2.45 (1H, m), 2.13 (1H, m), 1.38 (4.5H, bs), 1.27
(4.5H, bs). Exact mass (FAB, M+H.sup.+) calcd for C.sub.18 H.sub.25
N.sub.2 O.sub.5 : 349.1763; Found: 349.1761.
EXAMPLE 9
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepin-1-yl]
acetyl]-L-methionine (9) ##STR2984##
The compounds synthesized via the route shown in Scheme 2 followed
standard solid-phase methodologies (Barany, G. and Merrifield, R.
B. (1980) in "The Peptides", 2, 1-284. Gross, E. and Meienhofer, J.
Eds. Academic Press, New York).
3-(tert-Butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benz-azapin-2
-one-1-acetic acid (1.6 mmol, 558 mg),
benzotriazol-1-yloxy-tris-(dimethylamino)-phosphonium
hexafluorophosphate (BOP, 1.6 mmol, 706 mg), N-methylmorpholine
(NMM, 1.6 mmol, 217 ul), and N-hydroxbenztriazole (HOBt, 1.6 mmol,
175 mg) in dimethylacetamide (DMA, 30 ml) were added to deprotected
L-methionine-linked Merrifield resin (Bachem, 1.5 gm, 0.71 meq/gm,
12hrs.). After wash (DMA, then dichloromethane (DCM)) and
deprotection steps (45% TFA/5% anisole/5% EtSMe/DCM), the resin was
neutralized (20% Et.sub.3 N/DCM) and washed (DCM). Next,
Fmoc-(S-trityl)-L-cysteine (4.3 mmol, 2.5 gm),
bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl, 4.7 mmol, 1.2
gm), and diisopropylethylamine (9.4 mmol, 1.6 ml) were combined and
added to the resin (DCM, 30 ml, 10 hrs). After removal of the Fmoc
(20% piperidine/DMA) and trityl (45% TFA/5% EtSMe/5% anisole/DCM)
protecting groups the resin was washed with MeOH, dried under
vacuum, cleaved from the resin (32 ml, HF/10% anisole/5% EtMeS,
0.degree. C., 1 hr.) and purified via HPLC. Purification of 119 mg
of crude material (Vydac C18, ACN/water/0.1% TFA) afforded the
product, N-[[3-(2-amino-3-mercapto-1-oxopropyl)
methylamino]-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepin-1-yl]
acetyl]-L-methionine, as two separable diastereomers (opposite
configuration at C-3 of the benzodiazepine) designated 9A (24 mg)
and 9B (27 mg) corresponding to the early and late eluting peaks
respectively.
Mass (electrospray, M+H.sup.+) calc: 483.1 found: 482.8 (9A), 482.8
(9B).
EXAMPLE 10
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)
amino]-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepin-1-yl]
acetyl]-L-methionine (10) ##STR2985##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-
1-acetic acid (428 mg, 1.3 mmol) was coupled to L-methionine resin
(1.2 gms, 0.71 mmol/gm) with BOP (565 mg, 1.3 mmol), NMM (170 ul,
1.3 mmol), and HOBt (140 mg, 1.3 mmol). Again, after deprotection
and washing steps, Fmoc-L-(S-trityl) cysteine (2.5 gm, 4.7 mmol)
was coupled using BOP-Cl (1.2 gm, 4.7 mmol) and DIPEA (1.6 ml, 9.4
mmol). Purification of 102 mg of the crude material yielded the two
diastereomers 10A (18 mg) and 10B (12 mg).
Mass (electrospray, M+H.sup.+) calc: 469.1 found: 468.8 (10A),
468.8 (10B).
In Examples 11-17 the parenthetical compound numbers refer to the
numbers in Scheme 3 below. ##STR2986##
EXAMPLE 11
3-Amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one (11)
To a suspension of 10.0 g (49.4 mmol) of
3-azido-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one (Example 3
above) in 100 mL of methanol/ethanol (1:1) was added 1 g of 10%
palladium on charcoal and the flask was purged with nitrogen and
charged with 55 psig hydrogen. The mixture was shaken for 5 h,
filtered through celite and concentrated. The residue was
recrystallized from MeOH to give 6.77 g (78%) in two crops. .sup.1
H NMR (300 MHz, CDCl.sub.3) d 8.38 (1H, bs), 7.3-7.1 (3H, m), 6.98
(1H, d, J=8 Hz), 3.42 (1H, dd, J=11, 7 Hz), 2.90 (1H, m), 2.70-2.40
(2H, m), 1.89 (1H, m), 1.78 (bs, 2H).
EXAMPLE 12
3(R)-Amino-2,3,4,5-tetrahydro-1H-[1] -benzazapin-2-one (12)
A suspension of 6.75 g (38.3 mmol) of
3-amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one and 5.18 g (34.5
mmol) of D-tartaric acid in 80 ml of ethanol/water (4:1) was warmed
to effect solution and aged overnight at ambient temperature. The
crystals were collected, recrystallized again from ethanol/water,
and dried under vacuum to give 4.61g (37%) of purified D-tartrate
salt. The salt was dissolved in water, solid potassium carbonate
was added until the pH was 10-11, and the solution extracted four
times with methylene chloride. The combined organics were dried
over magnesium sulfate, filtered, and concentrated in vacuo to give
2.03 g (30% overall) of the 3(R)-amine. [.alpha.]D=+407.degree.
(c=1, MeOH); lit. Fisher, M. H.; et al EP 513974-A1, 2/28/92,
[.alpha.]D=+455.degree. (c=1, MeOH).
EXAMPLE 13
3(S)-Amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one (13)
The mother liquors from Example 12, enriched in the 3(S)-isomer
were free-based as above, treated with L-tartaric acid (1 eq), and
recrystallized from ethanol/water. The crystals were collected,
free-based with aqueous potassium carbonate and extracted as above.
Drying and concentration gave 1.50 g (22% overall) of the
3(S)-amine.
EXAMPLE 14
Ethyl
3(S)-3-amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-acetate
(14)
To a solution of 1.50 g (8.51 mmol) of
3(S)-amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one in 20 mL of
dimethylformamide at 0.degree. C. was added 340 mg (8.51 mmol, 60%
disp. in mineral oil) of sodium hydride. The suspension was warmed
to ambient temperature for 1.5 h, recooled to 0.degree. C., and
treated with 1.42 g (0.948 mL, 8.51 mmol) of ethyl bromoacetate in
5 mL of dimethylformamide. The ice bath was removed, and after 1 h
the mixture was concentrated in vacuo. The residue was partitioned
between ethyl acetate and 1N sodium bicarbonate, the organic phase
removed, washed with brine, dried over magnesium sulfate, and
concentrated. Trituration with ether gave 767 mg (34%) of a
colorless solid in the first crop. [.alpha.]D=-295.degree. (c=0.95,
EtOH); (lit. Watthey, J. W. H.; et al J. Med. Chem. 1985, 28,
1511., [.alpha.]D= -285.5.degree. (c=0.99, EtOH)). .sup.1 H NMR
(300 MHz, CDCl.sub.3) d 7.30-7.10 (4H, m), 4.62 (1H, d, J=17.8 Hz),
4.45 (1H, d, J=17.8 Hz), 4.19 (2H, q, J=7 Hz), 3.43 (1H, m), 3.23
(1H, m), 2.58 (1H, m), 2.41 (1H, m), 1.78 (2H, bs), 1.26 (3H, t,
J=7 Hz).
EXAMPLE 15
Ethyl
3(S)-3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazepin-2-o
ne-1-acetate (15)
Di-t-butyl-dicarbonate (1.23 g, 5.62 mmol) was added to a slurry of
737 mg (2.81 mmol) of Ethyl
3(S)-3-amino-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-one-1-acetate
in 20 mL methylene chloride/water (1:1) and the mixture was rapidly
stirred at ambient temperature for 30 min. The organic phase was
separated and washed with 1N sodium bicarbonate, brine, dried over
magnesium sulfate, and filtered. Concentration in vacuo gave a foam
that was chromatographed (70 g silica gel 60, ethyl acetate/hexane
(1:2)) to give 987 mg (97%) of a colorless foam identical with
material prepared above (Example 5).
EXAMPLE 16
Ethyl
3(S)-3-(tert-butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazap
in-2-one-1-acetate (16)
To a 0.degree. C. solution of 488 mg (1.35 mmol) of Ethyl
3(S)-3-(tert-butoxycarbonylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazapin-2-o
ne-1-acetate in 5 mL of dimethylformamide was added 0.170 mL (2.69
mmol) of methyl iodide and 59 mg of sodium hydride (60% dispersion
in mineral oil, 1.48 mmol). The suspension was stirred at ambient
temperature for 15 h and partitioned between ethyl acetate and 1N
sodium bicarbonate. The organic phase was washed with 3% sodium
bisulfite, water, brine, dried over magnesium sulfate, filtered,
and concentrated. Chromatography (35 g silica gel 60, ethyl
acetate/hexane (1:3)) gave 410 mg (81%) of a colorless foam. .sup.1
H NMR (300 MHz, CDCl.sub.3, spectrum broad due to carbamate
rotomers) d 7.3-7.1 (4H, m), 4.90-4.05 (5H, bm), 3.35 (1H, bm),
3.01 (3H, s), 2.65 (1H, m), 2.48 (1H, m), 2.15 (1H, m), 1.4-1.1
(12H, b).
EXAMPLE 17
(S)-3-(tert-Butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazepin
-2-one-1-acetic acid (17)
To a 0.degree. C. solution of 410 mg (1.09 mmol) of Ethyl
3(S)-3-(tert-butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazap
in-2-one-1-acetate in 6 mL of tetrahydrofuran/water (2:1) was added
2.2 mL of 1N sodium hydroxide. The mixture was magnetically stirred
for 3 h at ambient temperature and concentrated in vacuo to remove
the volatiles. The slurry was diluted with water, extracted with
ether (discarded), and acidified to pH 2 with 1N sodium hydrogen
sulfate. The aqueous phase was extracted twice with ethyl acetate
and the combined organics were washed with brine, dried over
magnesium sulfate, filtered, and concentrated to a colorless foam.
This material was identical with racemic material prepared above
(Example 8). [.alpha.]D=-177.degree. (c=0.90, EtOH).
EXAMPLE 18
N-[[3(S)-3-(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepin-1-yl]acetyl]-L-methi
onine (18) ##STR2987##
The title compound was prepared using the procedure of Example 9 in
which
3(S)-3-(tert-butoxycarbonylmethylamino)-2,3,4,5-tetrahydro-1H-[1]-benzazep
in-2-one-1-acetic acid (338 mg, 0.8 mmol) was coupled to
L-methionine resin (0.75 gm, 0.71 mmol/gm) using BOP (353 mg, 0.8
mmol), NMM (108 ul, 0.8 mmol), and HOBt (88 mg, 0.8 mmol). Again,
after deprotection and washing steps, Fmoc-L-(S-trityl) cysteine
(2.5 gm, 4.7 mmol) was coupled using BOP-Cl (1.2 gm, 4.7 mmol) and
DIPEA (1.6 ml, 9.4 mmol). HPLC analysis of the product showed that
it was identical to a single diastereomer of the compound shown in
Example 9. Co-injection of the compound prepared in Example 18 with
the diastereomeric mixture of compounds prepared in Example 9
showed that the material prepared in Example 18 co-eluted with the
first peak (9A) of Example 9. Therefore, for these compounds and
all other molecules of similar structure described here we can
conclude the configuration at the 3-carbon of the seven-membered
ring is such that the 3(S) isomer elutes first (A peak) and the
3(R) isomer follows (B peak). It should be noted that in the
benzodiazepine series the assignment of stereochemistry reverses
from that of the benzazepine compound discussed in this example due
to a shift in priority of the atoms bound to C-3 of the
seven-membered ring. Therefore, in the benzodiazepine examples
discussed below, we can conclude that the 3(R) isomer elutes first,
and the 3(S) isomer follows.
In Examples 19-26 the parenthetical compound numbers refer to the
numbers in Scheme 4 below. ##STR2988##
EXAMPLE 19
2-Bromoacetamido-benzophenone (19)
A solution of bromoacetyl bromide (100 mL, 1.15 mol, Aldrich)
dissolved in dichloromethane (300 mL) was added over 30 min to a
solution of 2-aminobenzophenone (197 g, 1.0 mol, Fluka) dissolved
in dichloromethane (1.3 L) and water (100) mL) cooled to
-10.degree. C. under vigorous mechanical stirring. The resulting
mixture was stirred for an additional 1 h at -5.degree. C. and then
was allowed to warm to ambient temperature. The layers were
separated, and the organic extract was washed with dilute sodium
bicarbonate, then was dried over sodium sulfate. Evaporation
afforded 309.8 g (95%) of 2-bromoacetamidobenzophenone as off-white
crystals.
EXAMPLE 20
2,3-Dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one (20)
A suspension of 2-bromoacetamidobenzophenone (275 g, 0.86 mol) in
methanol (1 L) was treated with a solution of saturated ammonia in
methanol (3 L), and the resulting solution was stirred at ambient
temperature for 6 h, then was heated at reflux for an additional 4
h. After cooling, water (500 mL) was added, and the solution was
concentrated by evaporation to about 1 L in volume, yielding
crystalline 2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one (20)
(200.7 g, 98%).
EXAMPLE 21
tert-Butyl-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetate
(21)
A 1 L round-bottomed flask was equipped with a magnetic stirring
bar and nitrogen inlet and was sequentially charged with 100 g
(0.423 mol) of 2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one, 600
mL of 1-methyl-2-pyrrolidinone (Aldrich, anhydrous), 97 mL (117 g,
0.601 mol) of tert-butyl bromoacetate (Aldrich), and 194 g (0.595
mol) of cesium carbonate (Aldrich). After stirring overnight at
room temperature, the reaction mixture was diluted with 2 L H.sub.2
O and extracted with EtOAc (3.times.600 mL). The combined organic
extracts were washed with H.sub.2 O (4.times.300 mL) and brine (200
mL), dried with anhydrous Na.sub.2 SO.sub.4, filtered, and
concentrated under reduced pressure to provide 202 g of a solid.
This material was recrystallized from hexanes/EtOAc to provide 123
g (83%) of
tert-butyl-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetate
(21) as a white crystalline solid.
EXAMPLE 22
2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic acid
(22)
A solution of
tert-butyl-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetate
(58 g, 0.172 mol) in neat trifluoroacetic acid (100 mL) was stirred
overnight, followed by evaporation and retreatment with an
additional amount of TFA (100 mL). The mixture was evaporated, and
the residue was dissolved in dichloromethane, and was washed with
water and brine. The organic layer was dried over sodium sulfate
and evaporated to yield
2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic acid (22)
(48.4 g, 100%) as a yellow foam.
EXAMPLE 23
3-oximino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (23)
A solution of
2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic acid (22)
(30 g, 0.106 mol) in glyme (1 L) was cooled to -5.degree. C. and
degassed under nitrogen. Solid potassium tert-butoxide (47.7 g,
0.43 mol) was added portionwise, and the resulting red solution was
stirred for 30 min at 0.degree.-5.degree. C. A solution of isobutyl
nitrite (13.8 mL, 0.117 mol, Aldrich) in glyme (20 mL) was then
added, producing an orange-yellow solid. The mixture was
neutralized with acetic acid (200 mL) with slight warming, and then
was evaporated. The residue was partitioned between butanol and
brine, and the organic layer was dried over sodium sulfate.
Additional residual inorganics precipitated on standing, and were
removed by filtration. Addition of hexane to the butanol solution
afforded a crude precipitate (34.3 g), which was recrystallized
from ethyl acetate-ethanol to yield
3-oximino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (23) (21.85 g, 66%).
EXAMPLE 24
3-Amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (24)
A solution of
3-oximino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (15.0 g, 48 mmol) in methanol (1 L) was hydrogenated over
catalytic ruthenium on carbon (5.0 g, Aldrich) at 40 psi and
70.degree. C. for 4 days. The catalyst was removed by filtration,
and the solution was evaporated to yield a crude solid (13.5 g).
Flash chromatography (50 ethyl acetate: 49 methanol: 1 water)
yielded pure
3-amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (9.69 g).
Alternatively, a solution of
3-oximino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (33 g, 102 mmol) in methanol (200 ml) containing 2 ml acetic
acid, was hydrogenated over Raney Nickel (1:1 by weight to oxime,
washed twice with water than once with ethanol) at 65 psi and 70 C
for 11/2 days. The catalyst was removed by suction filtration
through celite, and the solution evaporated to yield crude
3-amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid which was used directly in the next step (see alternative
synthesis in Example 25 below).
EXAMPLE 25
3-(tert-Butoxycarbonyl)amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-on
e-1-acetic acid (25)
A solution of
3-amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid (1.0 g, 3.37 mmol) in DMF (10 mL) and water (5 mL) was treated
with triethylamine (0.34 g, 3.37 mmol) and di-tert-butyl
dicarbonate (0.73 g, 3.37 mmol) under nitrogen. After stirring at
ambient temperature overnight, the mixture was evaporated and the
residue was partitioned between ethyl acetate and water (pH=2 w/6N
HCl). The organic extract was dried over sodium sulfate and
evaporated. Flash chromatography (80 ethyl acetate: 19 methanol: 1
water) of the residue afforded
3-(tert-butoxycarbonyl)amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-o
ne-1-acetic acid (940 mg) as a yellow solid.
.sup.1 HNMR: consistent with structure
Mass (electrospray, M+H.sup.+) calc: 410.2 found: 410.0.
Alternatively,
3-amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-one-1-acetic
acid prepared above (alternative synthesis, Example 24) in THF (100
ml) and water (100 ml) was cooled to 0 C and di-tert-butyl
dicarbonate (28.8 g, 132 mmol) was added under nitrogen followed by
1N NaOH until the pH of the solution was .about.10. The solution
was allowed to come to ambient temperature, stirred overnight, and
cooled again to 0.degree. C. and acidified (pH .about.3) with
dropwise addition of concentrated H2SO4. The solution was
partitioned (EtOAc) and the organic extract dried over sodium
sulfate and evaporated. The residue was recrystallized from 120 ml
MeOH yielding 18 g (44 mmol, 37%) of
3-(tert-butoxycarbonyl)amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-o
ne-1-acetic acid as yellow needles.
EXAMPLE 26
3-(tert-Butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepi
n-2-one-1-acetic acid (26)
An oven-dried, 100 mL round-bottomed flask was equipped with a
magnetic stirring bar and nitrogen inlet and was sequentially
charged with 4.628 g (11.3 mmol) of
3-(tert-butoxycarbonyl)amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-o
ne-1-acetic acid, 50 mL of anhydrous THF (Aldrich), and 2.80 mL
(6.38 g, 45.0 mmol) of methyl iodide (Aldrich). The reaction flask
was cooled to -5.degree. C. in an ice/acetone bath and 1.18 g (29.5
mmol) of a 60% oil dispersion of sodium hydride was added in
portions over a 5 min period (caution: vigorous gas evolution).
After a 50-min period, the reaction mixture was quenched with a 5%
(w/v) aqueous solution of citric acid, diluted with H.sub.2 O and
extracted with EtOAc (3.times.40 mL). The combined organics were
washed with H.sub.2 O (30 mL), brine (30 mL), dried with anhydrous
Na.sub.2 SO.sub.4, filtered, and concentrated under reduced
pressure to provide 6.27 g of a viscous yellow oil. Flash
chromatography of the crude material on 150 g of silica using
43:55:2 EtOAc/hexanes/AcOH as eluent yielded 4.54 g (10.7 mmol,
95%) of
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid as a clear glass.
1HNMR consistent with structure.
Mass calc for C23H26N3O5: 424.1872 found: 424.1909
EXAMPLE 27
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (27)
##STR2989##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (675 mg, 1.6 mmol) was coupled to
L-methionine resin (1.5 gms, 0.71 mmol/gm) with BOP (710 mg, 1.6
mmol), HOBt (220 mg, 1.6 mmol), and NMM (180 ul, 1.6 mmol). Again,
after deprotection and washing steps, Fmoc-L-(S-trityl) cysteine
(2.5 gm, 4.7 mmol) was coupled using BOP-Cl (1.2 gm, 4.7 mmol) and
DIPEA (1.6 ml, 9.4 mmol). Purification of 107 mg of the crude
material yielded the two diastereomers 27A (27 mg) and 27B (15
mg).
Mass (FAB, M+H.sup.+) calc: 558.2 found:558.3 (27A), 558.3
(27B).
EXAMPLE 28
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)amino]-2,3-dihydro-2-oxo-5-phenyl-1H
-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (28) ##STR2990##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)amino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepin-2-o
ne-1-acetic acid (1.3 gm, 3.2 mmol) was coupled to L-methionine
resin (1.5 gms, 0.71 mmol/gm) with BOP (1.4 gm, 3.2 mmol), HOBt
(430 mg, 1.6 mmol), and NMM (350 ul, 3.2 mmol). Again, after
deprotection and washing steps, Fmoc-L-(S-trityl) cysteine (2.5 gm,
4.7 mmol) was coupled using BOP (1.9 gm, 4.3 mmol), HOBt (600 mg,
4.3 mmol), and NMM (500 .mu.l, 4.3 mmol). Purification of 107 mg of
the crude material yielded the two diastereomers 28A (38 mg) and
28B (14 mg).
Mass (electrospray, M+H.sup.+) calc: 544.2 found:544.8 (28A), 544.8
(28B).
EXAMPLE 29
N-[[3-[(3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1
,4-benzodiazepin-1-yl]acetyl]-L-methionine (28) ##STR2991##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (0.45 gm, 1.1 mmol) was coupled to
L-methionine resin (1.0 gms, 0.71 mmol/gm) with BOP (0.47 gm, 1.1
mmol), HOBt (150 mg, 1.1 mmol), and NMM (120 ul, 1.1 mmol). Again,
after deprotection and washing steps, (S-methylbenzyl)propionic
acid (0.6 gm, 2.8 mmol) was coupled using BOP-Cl (0.8 gm, 3.1 mmol)
and DIPEA (1.09 ml, 6.2 mmol). Purification of 150 mg of the crude
material yielded the two diastereomers as an unseparable mixture
29A+B (9.5 mg).
Mass (electrospray, M+H.sup.+) calc: 543.2 found:543.3 (29A+B).
EXAMPLE 30
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)amino]-2,3-dihydro-2-oxo-5-phenyl-1H
-1,4-benzodiazepin-1-yl]acetyl]-L-serine (30) ##STR2992##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (0.47 gm, 1.1 mmol) was coupled to
L-(O-benzyl)serine resin (0.8 gms, 0.92 mmol/gm) with BOP (0.49 gm,
1.1 mmol), HOBt (150 mg, 1.1 mmol), and NMM (120 .mu.l, 1.1 mmol).
Again, after deprotection and washing steps, Fmoc-L-(S-trityl)
cysteine (1.65 gm, 2.8 mmol) was coupled using BOP-Cl (0.8 gm, 3.1
mmol) and DIPEA (1.09 ml, 6.2 mmol). Purification of 160 mg of the
crude material yielded the two diastereomers 30A (14 mg) and 30B
(12 mg).
Mass (electrospray, M+H.sup.+) calc: 514.1 found:513.5 (30A), 513.5
(30B).
EXAMPLE 31
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-leucine (31)
##STR2993##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (0.45 gm, 1.1 mmol) was coupled to L-leucine
resin (1.5 gms, 0.47 mmol/gm) with BOP (0.47 gm, 1.1 mmol), HOBt
(140 mg, 1.1 mmol), and NMM (120 ul, 1.1 mmol). Again, after
deprotection and washing steps, Fmoc-L-(S-trityl) cysteine (1.65
gm, 2.8 mmol) was coupled using BOP-Cl (0.8 gm, 3.1 mmol) and DIPEA
(1.09 ml, 6.2 mmol). Purification of 86 mg of the crude material
yielded the two diastereomers 31A (16 mg) and 31B (15 mg).
Mass (electrospray, M+H.sup.+) calc: 540.2 found:540.3 (31A), 540.3
(31B).
EXAMPLE 32
N-[[3-[(2-acetylamino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo
-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (32)
##STR2994##
The title compound was prepared using the procedure of Example 9 in
which the synthesis of Example 27 was conducted, and the
amino-terminus acetylated via treatment with acetic anhydride (5%)
in 5% NMM/DCM for 5 min. Purification of 204 mg of the crude
material yielded the two diastereomers 32A (30 mg) and 32B (36
mg).
Mass (electrospray, M-H.sup.+) calc: 598.2 found:598.1 (32A), 598.1
(32B).
EXAMPLE 33
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine amide (33)
##STR2995##
The title compound was prepared using the procedure of Example 9 in
which Boc-L-methionine (0.6 gm, 2.4 mmol) was coupled to MBHA-resin
(1.5 gm, 0.53 mmol/gm) using BOP (0.33 gm, 2.4 mmol), HOBt (0.26
gm, 2.4 mmol), and NMM (130 .mu.l, 2.4 mmol). Following suitable
deprotection and washing steps,
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (0.5 mg, 1.2 mmol) was coupled with BOP
(0.53 mg, 1.2 mmol), HOBt (160 mg, 1.2 mmol), and NMM (130 .mu.l,
1.2 mmol). Again, after deprotection and washing steps,
Fmoc-L-(S-trityl) cysteine (2.5 gm, 4.7 mmol) was coupled using
BOP-Cl (1.2 gm, 4.7 mmol) and DIPEA (1.6 ml, 9.4 mmol).
Purification of 300 mg of the crude material yielded the two
diastereomers 33A (43 mg) and 33B (42 mg).
Mass (electrospray, M-H.sup.+) calc: 557.2 found:557.9 (33A), 556.9
(33B).
EXAMPLE 34
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-phenylalanine (34)
##STR2996##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid was coupled to L-phenylalanine resin. Again,
after deprotection and washing steps, Fmoc-L-(S-trityl) cysteine
was coupled as above. Purification of 106 mg of the crude material
yielded the two diastereomers 34A (39 mg) and 34B (32 mg).
Mass (electrospray, M+H.sup.+) calc: 574.2 found:574.3 (34A), 574.3
(34B).
EXAMPLE 35
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-alanine (35)
##STR2997##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid was coupled to L-alanine resin. Again, after
deprotection and washing steps, Fmoc-L-(S-trityl) cysteine was
coupled as above. Purification of 116 mg of the crude material
yielded the two diastereomers 35A (26 mg) and 35B (28 mg).
Mass (electrospray, M+H.sup.+) calc: 498.2 found: 498 (35A), 498
(35B).
EXAMPLE 36
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-proline (36)
##STR2998##
The title compound was prepared using the procedure of Example 9 in
which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid was coupled to L-alanine resin. Again, after
deprotection and washing steps, Fmoc-L-(S-trityl) cysteine was
coupled as above. Purification of 114 mg of the crude material
yielded the two diastereomers 36A (28 mg) and 36B (27 mg).
Mass (electrospray, M+H.sup.+) calc: 524.2 found: 524.3 (36A),
524.3 (36B).
EXAMPLE 37
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]
acetyl]-L-methionine methyl ester (37) ##STR2999##
3-(tert-Butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepi
n-2-one-1-acetic acid (5.5 mmol, 2.3 gm), L-methionine methyl ester
(16.5 mmol, 2.7 gm, Sigma), diisopropylcarbodiimide (DIPC, 6.6
mmol, 1.04 ml), and HOBt (6.6 mmol, 0.9 gm) were combined in DCM
(20 ml). After 10 hrs, the reaction was diluted (DCM, 90 ml),
extracted (0.1N H.sub.2 SO.sub.4 then brine), dried (MgSO.sub.4),
and concentrated to yield 3.5 gm (113%) of crude
N-[[3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-2-oxo-5-phenyl-1H-1,
4-benzodiazepin-1-yl] acetyl]-L-methionine methyl ester.
0.75 gm of this material was deprotected (30% TFA, 30 ml, 3 hrs),
concentrated, neutralized via extraction in 100 ml EtOAc (saturated
NaHCO.sub.3 then brine), and purified (silica, DCM/MeOH/Et.sub.3 N
99:1:0.2%) to yield N-[[3-amino-2,3-dihydro-2-oxo-5-phenyl-1H-1,
4-benzodiazepin-1-yl] acetyl]-L-methionine methyl ester as a clear
oil (0.55 gm, 89%).
Reaction of this material with Boc-(S-ethylthio)-cysteine (3.6
mmol, 0.99 gm), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (EDC, 3.6 mmol, 0.68 gm), and HOBt (1.2 mmol, 0.16
gm) in DMF (10 ml, 12 hrs) was followed by concentration, aqueous
workup and chromatography (as above), to yield
N-[[3-[(2-(tert-butoxycarbonyl)amino-3-mercapto-1-oxopropyl)methylamino]-2
,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine
methyl ester (0.82 gm, 93%). After removal of the Boc-(30% TFA as
above) and ethylthio- (60 mg dithiothreitol, 20 ml 50% ACN/H.sub.2
O, pH 7.5) protecting groups,
N-[[3-(2-amino-3-mercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1, 4-benzodiazepin-
1-yl] acetyl]-L-methionine methyl ester was purified by HPLC (Vydac
C18, ACN/H.sub.2 O/TFA) which resolved each of diastereomers
possessing opposite configuration at C-3. These isomers were
designated A and B as before (see Example 9). Purification of 120
mg of the crude material yielded the two diastereomers 37A (26 mg)
and 37B (30 mg).
Mass (electrospray, M+H.sup.+) calc: 572.2 found: 572.3 (37A),
572.3 (37B).
EXAMPLE 38
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1, 4-benzodiazepin-1-yl]
acetyl]-L-methionine methyl ester (38) ##STR3000##
The title compound was prepared using the procedure of Example 37
in which an identical procedure was performed, omitting the step
for removal of the ethylthio protecting group. Purification of 140
mg of the crude material yielded the two diastereomers 38A (30 mg)
and 38B (30 mg).
Mass (electrospray, M+H.sup.+) calc: 632.3 found: 631.9 (38A),
631.9 (38B).
EXAMPLE 39
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1, 4-benzodiazepin-1-yl]
acetyl]-L-methionine ethyl ester (39) ##STR3001##
The title compound was prepared using the procedure of Example 37
in which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (0.85 mmol, 0.36 gm), L-methionine ethyl
ester (2.55 mmol, 0.45 gm), DIPC (1.02 mmol, 0.16 ml), and HOBt
(1.02 mmol, 0.14 gm) were combined in the first step. Subsequent
reactions to yield the title compound were completed in a manner
similar to that shown in Example 37. Purification of 150 mg of the
crude material yielded the two diastereomers 39A (21 mg) and 39B
(22 mg).
Mass (electrospray, M+H.sup.+) calc: 586.3 found: 585.9 (39A),
585.9 (39B).
EXAMPLE 40
N-[[3-(2-Amino-3-ethylthiomercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]
acetyl]-L-methionine ethyl ester (40) ##STR3002##
The title compound was prepared using the procedure of Example 37
in which an identical procedure was performed, omitting the step
for removal of the ethylthio protecting group. Purification of 150
mg of the crude material yielded the two diastereomers 40A and
40B.
Mass (electrospray, M+H.sup.+) calc: 646.3 found: 645.7 (40A),
645.7 (40B).
EXAMPLE 41
N-[[3-(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]
acetyl]-L-methionine cyclohexyl ester (41) ##STR3003##
L-methionine cyclohexyl ester was prepared by combining
Boc-L-methionine (2.5 g, 10 mmol), cyclohexanol (3.12 ml, 30 mmol),
DIPC (1.9 ml, 12 mmol) and 4-dimethylaminopyridine (DMAP, 0.12 g,
1.0 mmol) in DCM, followed by aqueous workup, flash chromatography
(silica, hexane/EtOAc (4:1)), removal of the Boc protecting group
(4N HCl/dioxane, 2 h), and basic workup (EtOAc/sat. Na.sub.2
CO.sub.3).
The title compound was prepared using the procedure of Example 37
in which
3-(tert-butoxycarbonyl)methylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazep
in-2-one-1-acetic acid (3.3 mmol, 1.4 gm), L-methionine cyclohexyl
ester (6.6 mmol, 1.53 gm), DIPC (3.96 mmol, 0.62 ml), and HOBt
(3.96 mmol, 0.54 gm) were combined in the first step. Subsequent
reactions to yield the title compound were completed in a manner
similar to that shown in Example 37. Purification of 150 mg of the
crude material yielded the two diastereomers 41A (35 mg) and 41B
(37 mg).
Mass (electrospray, M+H.sup.+) calc: 640.3 found: 640.1 (41A),
640.1 (41B).
The following examples were prepared similarly to above
syntheses.
EXAMPLE 42
N-[[3-[(2-ethylamino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-
5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (42)
##STR3004## The title compound was prepared as above with the
addition of a step following deprotection of the N-terminal
Boc-residue on cysteine. This step allowed attachment of an ethyl
moiety on nitrogen through reductive amination using acetaldehyde
and sodium cyanoborohydride in DMF/1% AcOH. The molecule was then
cleaved, purified and analyzed as above.
EXAMPLE 43
N-[[3-[(2-methylthiazolidine-4-carboxyl)methylamino]-2,3-dihydro-2-oxo-5-ph
enyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (43)
##STR3005## The title compound was prepared by coupling
N-Boc-2-methyl thiazolidine-4-carboxylic acid instead of cysteine.
Purified and analyzed as above.
EXAMPLE 44
N-[[3-[(2-Amino-3-mercapto-1-oxopropyl)
methylamino]-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-1-yl]acetyl]-
L-cysteine (44) ##STR3006## The title compound was prepared
following the protocol shown in Example 9, except
S-(4-methylbenzyl)-N-Boc-L-cysteine-linked Merrifield resin was
used.
EXAMPLE 45
N-[[3-[(2-amino-3-hydroxy-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phen
yl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine (45) ##STR3007##
The title compound was prepared following the protocol shown in
Example 9, except O-(tert-butyl)-N-Boc-L-serine was used in place
of cysteine.
EXAMPLE 46
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-leucine tetrazole (46)
##STR3008## The title compound was prepared using a similar
procedure to that shown for Example 37, except that tetrazole
derivative of leucine, denoted as L-leucine tetrazole, was used
instead of a carboxy-terminal ester. The synthesis of this material
is shown below in Scheme 6. ##STR3009## steps 1 and 2)
The N-Boc-L-leucine monohydrate (10 g, 40.2 mmol) and triethylamine
(5.6 mL, 40.2 mmol) were dissolved in 50 mL dry chloroform and
cooled to 0.degree. C. Ethyl chloroformate (3.84 mL, 40.2 mmol) was
add dropwise and stirred for 10 minutes. 40 mL dry chloroform
saturated with ammonia was added, stirred for 15 minutes, then
warmed to ambient temperature and stirred for 1 hr. Evaporation of
the solvent gave the crude product which was washed with aqueous
NaHCO.sub.3 then with water. The product was collected by
filtration, washed with water, and dried by vacuum to yield 8.53g
(37.1 mmol, 92% yield) of the desired amide as a white solid.
R.sub.f =0.36 (75%EtOAc/hexanes). .sup.1 H NMR (300MHz, CD.sub.3
OD): 4.05 (1H, br t), 1.70 (1H, m), 1.50 (2H, m), 1.45 (9H, s),
0.95 (6H, dd).
step 3)
The N-Boc-L-leucine amide (6.45g, 28 mmol) was dissolved in 40 mL
dry pyridine and cooled to -5.degree. C. Phosphorous oxychloride
(3.66 mL, 39.3 mmol), in 6 mL dry methylene chloride was added
dropwise and the solution was stirred at -5.degree. C. for 1 hr.
Ice water was added to the reaction and the nitrile was extracted
with EtOAc (3.times.200 mL). The combined organics were washed
successively with 100 mL water, 1N HCl (2.times.100 mL), aq.
NaHCO.sub.3 (2.times.100 mL) and 100 mL water. The organics were
dried over MgSO.sub.4 and concentrated to yield 4.27g (20.1 mmol,
72% yield) as a light yellow solid. R.sub.f =0.79
(75%EtOAc/hexanes). .sup.1 H NMR (300MHz, CD.sub.3 OD): 4.45 (1H,
br t), 1.75 (1H, m), 1.65 (2H, dd), 1.45 (9H, s), 0.95 (6H, dd).
IR: 2240 cm.sup.-1.
step 4)
The N-Boc-L-leucine nitrile (1.65g, 7.78 mmol), sodium azide
(0.53g, 8.15 mmol) and ammonium chloride (0.46g, 8.6 mmol) were
dissolved in 7 mL dry DMF and the flask was placed in a 105.degree.
C. oil bath. After 7 hr., sodium azide (0.26g, 4.0 mmol) and
ammonium chloride (0.27g, 5.0 mmol) were added and the solution was
stirred overnight at 105.degree. C. Cooled and concentrated. Flash
chromatography (7% MeOH in methylene chloride ramping to 25% MeOH
in methylene chloride) provided 1.93g (7.56 mmol, 97% yield) as a
white solid. R.sub.f =0.33 (10%MeOH/methylene chloride). .sup.1 H
NMR (300MHz, CD.sub.3 OD): 5.90 (1H, s), 5.15 (1H, br t), 1.60-1.90
(3H, m), 1.40 (9H, s), 0.95 (6H, dd).
preparation of the title compound:
The N-Boc-L-leucine tetrazole (0.45g, 1.75 mmol) was suspended in 5
mL 4N HCl/dioxane. Dissolves is 10 min. Concentrated after 2 hr. to
a tan foam. Dissolved in 5 mL water and taken to pH=9 with 1N NaOH.
The resulting free-amine was concentrated to a white solid and used
directly in the coupling reaction (below). R.sub.f =0.20
(25%MeOH/methylene chloride). .sup.1 H NMR (300MHz, CD.sub.3 OD):
4.40 (1H, br m), 3.65 (1H, m), 1.90 (1H, m), 1.75 (1H, m), 1.25
(1H, m) 0.95(6H, dd).
Coupling reactions were identical to those describe in Example 37.
The leucine tetrazole was suspended in 3 mL CH.sub.2 Cl.sub.2 -
does not dissolve.
3-(tert-Butoxycarbonylmethylamino-2,3-dihydro-5-phenyl-1H-1,4-benzodiazepi
n-2-one-1-acetic acid (0.343g, 0.81 mmol), HOBT (0.17g, 1.26 mmol)
and DIPC (1.1 mL, 1.1 mmol) were added and the solution was stirred
overnight. Workup and coupling of the cysteine was identical to the
procedure for Example 37. Purification of 50 mg of the crude
material yielded the two diastereomers 46A (7 mg) and 46B (8
mg).
Mass (electrospray, M+H.sup.+) calc: 564.2 found: 564.3 (46A),
564.3 (46B).
EXAMPLE 47
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]acetyl]-L-methionine tetrazole (47)
##STR3010## Synthesis of the title material was identical to that
describe in Example 46, substituting methionine for leucine.
Purification of 50 mg of the crude material yielded the two
diastereomers 47A (5 mg) and 47B (5 mg).
Mass (electrospray, M+H.sup.+) calc: 582.2 found: 582.4 (47A),
582.4 (47B).
EXAMPLE 48
N-[[3-[(2-amino-3-mercapto-1-oxopropyl)methylamino]-2,3-dihydro-2-oxo-5-phe
nyl-1H-1,4-benzodiazepin-1-yl]propionyl]-L-methionine (48)
##STR3011## The title compound was prepared using a procedure
similar to that described for Example 9, substituting the acetyl
modification at the N-1 of the benzodiazepine with a propionyl
moiety. This was accomplished using a modification of the protocol
used for the preparation of compound 26. Purification of 105 mg of
the crude material yielded the two diastereomers 48A (20 mg) and
48B (18 mg).
Mass (electrospray, M+H.sup.+) calc: 572.2 found: 573.1 (48A),
573.1 (48B).
EXAMPLE 49
In vitro inhibition of CAAX farnesyltransferase
All compounds were tested for in vitro inhibition of CAAX protein
farnesyltransferase. The enzyme was isolated and purified to
homogeneity from rat brain homogenates by sequential ammonium
sulfate fractionation, Mono Q ion-exchange chromatography, and
peptide affinity chromatography as described in Reiss, Y., Seabra,
M. C., Goldstein, J. L., and Brown, M. S. Methods: A Companion to
Methods in Enzymology 1, 241-245 (1990). Alternatively, recombinant
CAAX protein farnesyltransferase was also used in this assay.
Recombinant enzyme was produced in a baculovirus expression system
as in O'Reilly, D. R., Miller, L. K., and Luckow, V. A. Baculovirus
Expression Vectors: A Laboratory Manual (W. H. Freeman and Co., New
York, 1992) and in Reiss, Y., Brown, M. S., Goldstein, J. L., in
preparation. 72 hrs after infection, the cells were harvested and
disrupted and the enzyme isolated by chromatography on Q-Sepharose.
The recombinant farnesyltransferase was judged to be .about.90%
pure by Comassie blue staining after SDS gel electrophoresis.
In each experiment, varying concentrations of the inhibitor were
mixed with the enzyme, and the amount of [3H]farnesyl transferred
from all-trans-[3H]farnesyl pyrophosphate to recombinant p21H-ras
was measured in a filter binding assay as described in Reiss, Y.,
Goldstein, J. L., Seabra, M. C., Casey, P. J., and Brown, M. S.
Cell 62, 81-88 (1990). Briefly, the assay mixture contained, in a
final volume of 50 .mu.l, 50 mM Tris-chloride (pH 7.5), 50 .mu.M
ZnCl.sub.2, 3 mM MgCl.sub.2, 20 mM KCl, 5 mM dithiothreitol (DTT),
0.4% (v/v) octyl-.beta.-glucoside, 1% (v/v) dimethyl sulfoxide
(DMSO), 0.6 .mu.M all-trans-[3H]farnesyl pyrophosphate (9730
dpm/pmol; Dupont-New England Nuclear), 40 .mu.M recombinant
p21H-ras (see Reiss, Y., Goldstein, J. L., Seabra, M. C., Casey, P.
J., and Brown, M. S. Cell 62, 81-88 (1990)), 10 ng purified CAAX
farnesyltransferase and concentrations of the indicated inhibitor
(varying from 0.1 nM to 10 .mu.M). After incubation for 30 min at
37 C, the amount of [3H]farnesyl group transferred to p21H-ras was
measured by precipitation with SDS/trichloroacetic acid, filtration
onto nitrocellulose, and scintillation counting. Immediately before
use, each inhibitor to be tested was diluted into a solution
containing 2.5% DMSO, 10 mM DTT, and 0.5% octyl-.beta.-glucoside,
and added to the 50 .mu.l reaction mixture in a volume of 20 .mu.l.
EC.sub.50 values were obtained as the estimated concentration of
inhibitor yielding 50% of the [3H]-recovered in control samples (no
inhibitor).
Structures of the compounds tested are reproduced below. Compound
numbers also refer to example numbers above. Table D below shows
the results of the CAAX farnesyltransferase assay for each
diastereomer (A and B) (except where noted, compound 29).
TABLE A
__________________________________________________________________________
##STR3012## cpd. R.sup.8 R.sup.24 R.sup.25
__________________________________________________________________________
27 CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3 CH(CH.sub.2
SH)NH.sub.2 28 CH((CH.sub.2).sub.2 SCH.sub.3)COOH H CH(CH.sub.2
SH)NH.sub.2 29 CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3
(CH.sub.2).sub.2 SH 30 CH(CH.sub.2 OH)COOH CH.sub.3 CH(CH.sub.2
SH)NH.sub.2 31 CH(CH.sub.2 CH(CH.sub.3).sub.2)COOH CH.sub.3
CH(CH.sub.2 SH)NH.sub.2 32 CH((CH.sub.2).sub.2 SCH.sub.3)COOH
CH.sub.3 CH(CH.sub.2 SH)NHCOCH.sub.3 33 CH((CH.sub.2).sub.2
SCH.sub.3)CONH.sub.2 CH.sub.3 CH(CH.sub.2 SH)NH.sub.2 34
CH(CH.sub.2 C.sub.6 H.sub.5)COOH CH.sub.3 CH(CH.sub.2 SH)NH.sub.2
35 CH(CH.sub.3)COOH CH.sub.3 CH(CH.sub.2 SH)NH.sub.2 42
CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3 CH(CH.sub.2
SH)NHC.sub.2 H.sub.5 43 CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3
CH(CH.sub.2 SCH(CH.sub.3))NH 44 CH(CH.sub.2 SH)COOH CH.sub.3
CH(CH.sub.2 SH)NH.sub.2 45 CH((CH.sub.2).sub.2 SCH.sub.3)COOH
CH.sub.3 CH(CH.sub.2 OH)NH.sub.2
__________________________________________________________________________
TABLE B ______________________________________ ##STR3013## cpd.
R.sup.8 R.sup.24 R.sup.25 ______________________________________ 37
CH((CH.sub.2).sub.2 SCH.sub.3)CO.sub.2 CH.sub.3 CH.sub.3
CH(CH.sub.2 SH)NH.sub.2 39 CH((CH.sub.2).sub.2 SCH.sub.3)CO.sub.2
CH.sub.2 CH.sub.3 CH.sub.3 CH(CH.sub.2 SH)NH.sub.2 41
CH((CH.sub.2).sub.2 SCH.sub.3)CO.sub.2 C.sub.6 H.sub.11 CH.sub.3
CH(CH.sub.2 SH)NH.sub.2 46 CH(CH.sub.2 CH(CH.sub.3).sub.2)CN.sub.4
H CH.sub.3 CH(CH.sub.2 SH)NH.sub.2 47 CH((CH.sub.2).sub.2
SCH.sub.3)CN.sub.4 H CH.sub.3 CH(CH.sub.2 SH)NH.sub.2
______________________________________
TABLE C
__________________________________________________________________________
##STR3014## cpd. R.sup.8 R.sup.24 R.sup.25 R.sup.1 T.sub.1 -T.sub.2
Z
__________________________________________________________________________
9 CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3 CH(CH.sub.2
SH)NH.sub.2 H CHCH.sub.2 CH.sub.2 10 CH((CH.sub.2).sub.2
SCH.sub.3)COOH H CH(CH.sub.2 SH)NH.sub.2 H CHCH.sub.2 CH.sub.2 48
CH((CH.sub.2).sub.2 SCH.sub.3)COOH CH.sub.3 CH(CH.sub.2 SH)NH.sub.2
C.sub.6 H.sub.5 CN CH.sub.2 CH.sub.2
__________________________________________________________________________
TABLE D ______________________________________ In vitro Inhibition
of CAAX Famesyltransferase compound EC.sub.50 (.mu.M) "A" EC.sub.50
(.mu.M) "B" ______________________________________ 9 1.2 0.024 10
1.6 1.4 27 0.8 0.0003 28 0.38 0.431 29 (unseparable) 1.8 1.8 30
10.0 0.0084 31 0.084 0.0021 32 1.2 8.0 33 2.2 0.019 34 0.54 0.0005
35 >10 0.022 36 >10 0.048 37 0.19 0.05 39 8.0 0.11 41 4.2
0.074 42 0.58 0.018 43 0.32 0.006 44 3.0 0.0045 45 >10 >10 46
0.26 0.0016 47 0.44 0.0046 48 2.5 0.004
______________________________________
EXAMPLE 50
Differential inhibition of CAAX farnesyltransferase, CAAX GG
transferase, and Rab GG transferase
FIG. 1 compares the inhibitory activity of compound 27B (denoted as
BZA-2B in the figure) on the three known protein
prenyltransferases. The compound inhibited recombinant ras CAAX
farnesyltransferase by 50% at 0.26 nM, which was nearly 200-fold
lower than the inhibitory concentration for the tetrapeptide CVFM.
Two other prenyltransferases, both of which transfer 20-carbon
geranylgeranyl (GG) groups, have been identified in rat brain. One
of these, CAAX GG transferase, attaches GG groups to proteins that
terminate in CAAX sequences in which X is leucine. The other
enzyme, Rab GG transferase, recognizes a different class of
substrates that do not terminate in CAAX sequences. Compound 27B
(denoted as BZA-2B in FIG. 1) inhibited the CAAX GG transferase at
a concentration more than 100-fold higher than that required to
inhibit CAAX farnesyltransferase (IC.sub.0.5 =35 nM; FIG. 1B) and
even less active on the third enzyme (IC.sub.0.5 =5.5 .mu.M, FIG.
1C).
The conditions used to assay the inhibition of CAAX
farnesyltransferase are identical to those described above for the
in vitro assay described in Example 50. In FIG. 1B the assay
mixture contained, in a final volume of 50 .mu.l, 50 mM sodium
Hepes (pH 7.2), 5 mM MgCl.sub.2, 5 mM DTT, 0.3 mM Nonidet P-40,
0.2% octyl .beta.-glucoside, 1% DMSO, 0.5 .mu.M all trans
[3H]geranylgeranyl pyrophosphate (33,000 dpm/pmol; ARC, Inc.), 5
.mu.M recombinant p21.sup.H -rasCVLL, 6.3 .mu.g partially purified
CAAX GG transferase (see Seabra, M. C., Reiss, Y., Casey, P. J.,
Brown, M. S., and Goldstein, J. L. Cell 268, 4055 (1993)) and
varying concentrations of the indicated inhibitor. In FIG. 1C, the
assay mixture contained in a final volume of 50 .mu.l, 50 mM sodium
Hepes (pH 7.2), 5 mM MgCl.sub.2, 5 mM DTT, 0.3 mM Nonidet P-40,
0.2% octyl .beta.-D-glucoside, 1% DMSO, 0.5 .mu.M all trans
[3H]geranylgeranyl pyrophosphate (33,000 dpm/pmol; ARC, Inc.), 2
.mu.M recombinant Rab1A, 2 ng each of purified Components A and B
of Rab GG transferase (see Seabra, M. C., Goldstein, J. L., Sudhof,
T. C., and Brown, M. S. J. Biol. Chem. 267, 14497 (1992)) and
varying concentrations of the indicated inhibitor.
EXAMPLE 51
Differential inhibition of CAAX farnesyltransferase and
geranylgeranyltransferase
FIG. 2 shows that compound 27B (denoted as BZA-2B in the figure)
which contains a C-terminal methionine, and compound 31B (denoted
as BZA-4B in the figure) which contains leucine, were equally
potent in inhibiting CAAX farnesyltransferase (IC.sub.0.5 =0.5 nM)
(FIG. 2A). This result is surprising because leucine-terminated
peptides are much less effective than methionine-terminated
peptides in inhibiting farnesylation of p21H-ras. Apparently,
important binding determinants are defined by the cysteine and
benzodiazepine substituents. The CAAX geranylgeranyltransferase was
5-fold more sensitive to the leucine-terminated inhibitor (FIG.
2B).
Each assay was carried out as described in Examples 49 and 50.
EXAMPLE 52
Quantitation of Farnesyl Group Transfer
As shown for some peptides, several inhibitors tightly bind the
enzyme and inhibit its activity, yet are not a substrate for the
enzyme. The ability of each compound to act as a substrate for the
CAAX farnesyltransferase was examined directly using thin layer
chromatography (see Table E) as described in Goldstein, J. L.,
Brown, M. S., Stradley, S. J., Reiss, Y., and Gierasch, L. M. J.
Biol. Chem. 266, 15575-15578 (1991) and Brown, M. S., Goldstein, J.
L., Paris, K. J., Burnier, J. P., and Marsters, Jr., J. C. Proc.
Natl. Acad. Sci. USA 89, 8313 (1992). Briefly, each 25 ul reaction
mixture contained 50 mM Tris-chloride (pH 7.5), 50 .mu.M
ZnCl.sub.2, 3 mM MgCl.sub.2, 20 mM KCl, 1 mM dithiothreitol (DTT),
0.2% (v/v) octyl-.beta.-glucoside, either 0.6 or 2.4 .mu.M
all-trans-[3H]farnesyl pyrophosphate (44,000 dpm/pmol; Dupont-New
England Nuclear), .about.5 ng purified CAAX farnesyltransferase and
90 pmol of the inhibitor to be tested (3.6 .mu.M). After incubation
at 37.degree. C. for 30 min, the reaction was stopped by addition
of 2 .mu.l of 250 mM EDTA, and the entire reaction mixture was
spotted onto a plastic-backed Silica Gel G thin layer sheet
(20.times.20 cm, Brinkmann Inst.) and placed in a tank containing
n-propyl alcohol/ammonium hydroxide/water (6:3:1 v/v/v). The
chromatogram was run for 3 hr, after which it was either subjected
to autoradiography or quantified by scintillation counting (see
Brown, M. S., Goldstein, J. L., Paris, K. J., Burnier, J. P., and
Marsters, Jr., J. C. Proc. Natl. Acad. Sci. USA 89, 8313
(1992)).
TABLE E ______________________________________ In vitro
Farnesylation by CAAX Farnesyltransferase compound % farnesylation
.sup.a "A" % farnesylation .sup.a "B"
______________________________________ 9 <1 <1 10 20 <1 27
5 <1 28 9 <1 29 <1 <1 30 <1 <1 31 <1 <1 32
110 <1 34 <1 37 <1 2 43 <1 <1 46 <1
______________________________________ .sup.a % farnesylation
measured as a percentage of [3H]farnesyl transferred to the
compound relative to a separate experiment using a goo substrate,
the tetrapeptide CVIM.
EXAMPLE 53
Assay of Inhibition of CAAX Farnesyltransferase in Cultured
Cells.
To study farnesylation in intact cells we used Met18b-2 cells, a
line of Chinese hamster ovary (CHO) cells that takes up
[3H]mevalonate efficiently (see Faust, J., and Krieger, M. J. Biol.
Chem. 262, 1966 (1987)) owing to the production of a mevalonate
transport protein (see Kim, C. M., Goldstein, J. L., Brown, M. S.
J. Biol. Chem. 267, 23113 (1992)). The [3H]mevalonate is converted
by the cells into [3H]farnesyl pyrophosphate and [3H]geranylgeranyl
pyrophosphate, which are then attached to proteins (Kim, C. M.,
Goldstein, J. L., Brown, M. S. J. Biol. Chem. 267, 23113 (1992)).
In the assay, stock cultures of Met18b-2 cells were seeded at a
density of 3.times.105 cells per 60 mm dish in 3 ml of medium A
(Dulbecco's modified Eagle medium/Ham's F12 medium (1:1, v/v)
containing 100 U/ml penicillin and 100 .mu.g/ml streptomycin)
supplemented with 5% (v/v) fetal calf serum (FCS). On day 3, each
monolayer was refed with 1 ml medium A supplemented with 1% FCS
(dialyzed against 0.15M NaCl). At this point the cells were treated
with 100 .mu.M compactin (see Brown, M. S., Faust, J. R.,
Goldstein, J. L., Kaneko, I., and Endo, A. J. Biol. Chem. 253, 1121
(1978)) which blocks the synthesis of unlabeled mevalonate within
the cells. Each compound was dissolved at 25 mM in DMSO/10 mM DTT
immediately before use and 10 .mu.l of this solution added
directly. After 2 hr. incubation at 37.degree. C., each monolayer
received 100 .mu.Ci [3H]mevalonate (60 Ci/mmol, American
Radiolabeled Chemicals, Inc.) added in 100 .mu.l of the medium A,
and the incubation continued for 4 hr. The cells were harvested by
rinsing three times with 3 ml of 50 mM Tris-HCl/0.15M NaCl (pH
7.5). 300 .mu.l of lysis buffer (0.5 .times. Dulbecco's
phosphate-buffered saline containing 1% Triton X-100, 5 .mu.g/ml
leupeptin, 5 .mu.g/ml pepstatin, 0.5 mM phenylmethylsulfonyl
fluoride, and 0.05 U/ml aprotinin) was added to each monolayer.
After incubation on ice for 5 min., the lysates were centrifuged 30
seconds in a microfuge at 12,000 g. The resulting supernatant was
tranferred to a new tube, and each pellet was resuspended in 60
.mu.l of lysis buffer. Protein concentrations were determined using
the BCA protein assay reagent (Pierce) according to the
manufacturer's instructions. Detergent-soluble (supernatant) and
-insoluble (pellet) samples were mixed with 2.times. SDS sample
buffer (see Laemmli, U. K. Nature 227, 680 (1970)) and heated at
95.degree. C. for 5 min before electrophoresis. Each lane contained
90 .mu.g protein, and was run on a 12.5% SDS-polyacrylamide gel.
The gel was dried, treated with ENTENSIFY (NEN-DuPont), and exposed
to Kodak XOMAT-AR film for 9 h at -80.degree. C. to allow
visualization of all prenylated proteins. Migration of
[14C]methylated molecular weight standards (Amersham) were used as
markers.
The results of this assay are shown in FIG. 3, using a final
concentration of 250 .mu.M for each inhibitor. Pilot experiments
showed that the tetrapeptide SVIM which does not inhibit
prenyltransferases did not alter the pattern of farnesylated
proteins in the cells, and this was used as a control for all
experiments. The Triton-soluble proteins marked F1 and F2 (unknown
functions) have been shown previously to be farnesylated (see
Reese, J. H., and Maltese, W. A. Mol. Cell. Biochem. 104, 109
(1991) and James, G., Brown, M. S., and Goldstein, J. L.,
unpublished observations). Protein F3 has been recently purified in
the Brown and Goldstein laboratory, and was also demonstrated to be
farnesylated. The labeled bands in the 20 to 27-kDa range consist
largely of low molecular weight GTP binding proteins, the vast
majority of which are geranylgeranylated. The major proteins in the
Triton-insoluble pellet are the nuclear lamins A and B, which are
farnesylated (see Farnsworth, C. C., Wolda, S. L., Gelb, M. H., and
Glomset, J. A. J. Biol. Chem. 264, 20422 (1989)). As shown in the
figure, none of the control peptides or control
benzodiazepine-peptideomimetics altered the prenylation of proteins
in intact cells. The high affinity inhibitors 27B and 31B (denoted
as BZA-2B and BZA-4B in the figure respectively) markedly decreased
the labeling of all three Triton-soluble farnesylated proteins (F1
to F3) and reduced moderately the labeling of the farnesylated
lainins (F-NL). The inhibitors had little effect on the low
molecular weight GTP binding proteins (SMG).
EXAMPLE 54
Dose Dependence of Inhibition of CAAX Farnesyltransferase in
Cultured Cells
Using the same cells and procedures followed above for Example 53,
we measured the dose dependence upon the inhibition of
farnesylation in intact cells. Concentration curves for three of
the more potent inhibitors are shown in FIG. 4 for the
Triton-soluble proteins. The pro-drug, compound 37B (denoted as
BZA-5B in the figure), was much more potent than the parent,
compound 27B (denoted as BZA-2B in the figure). Compound 37B
inhibited the farnesylation of F1-3 detectably at 10 .mu.M, and
almost completely at 25 .mu.M. Compound 33B, the C-terminal amide,
was of intermediate potency.
EXAMPLE 55
Immunoprecipitation of [3H]-labeled ras Proteins
To demonstrate the inhibition of farnesylation of ras proteins
directly (see FIG. 5), Met18b-2 cells were incubated with
[3H]mevalonate and then immunoprecipitated the cell lysates with a
monoclonal antibody that reacts with all four ras proteins. In each
experiment, aliquots of the Triton X-100 fraction (300 .mu.g
protein, prepared as described above) were incubated with 1 .mu.g
anti-ras monoclonal antibody (Oncogene Sciences, Inc.) overnight at
4.degree. C. on a rotating platform. Immune complexes were
precipitated by addition of 25 .mu.l Protein A-agarose suspension
that had been pre-coated with goat anti-rat IgG (Oncogene Sciences,
Inc.) according to the manufacturer's instructions. After a 30 min
incubation at 4.degree. C., the agarose beads were pelleted by
centrifugation and washed 5 times with 1 ml each of wash solution
(50 mM Tris-HCl, 50 mM NaCl, 0.5% (w/v) deoxycholate, 0.5% (v/v)
Nonidet P-40, and 0.1% (w/v) SDS at pH 7.5). 75 .mu.l of 1 .times.
SDS sample buffer was added and each sample heated for 5 min at
95.degree. before electrophoresis as above. As seen in FIG. 5,
shows increasing the concentration of compound 37B (denoted as
BZA-5B in the figure) did not detectably inhibit the incorporation
of radioactivity into the abundant SMG proteins, most of which are
geranylgeranylated (lanes 2-4). However, at 50 .mu.M the compound
abolished incorporation of [3H] mevalonate into immunoprecipitated
ras proteins (lanes 7-9). Inhibition was readily detectable at 10
.mu.M. The control molecules, the tetrapeptide SVIM and compound
27A (denoted as BZA-2A in the figure), had no effect.
EXAMPLE 56
Assay of Changes in Morphology of ras-transformed Cells.
As shown in FIG. 6, rat-1 fibroblasts transformed with an activated
mutant of p21H-ras (Val 12) grow in multilayered clumps, indicative
of malignent transformation. Compound 27A (denoted as BZA-2A in the
figure) and compound 27B (denoted as BZA-2B in the figure) were
added at 250 .mu.M to test whether these compounds would induce
reversion to a more normal phenotype as had been previously shown
with microinjection of anti-ras antibodies (see Feramisco, J. R. et
al Nature 314, 639 (1985)). The H-ras transformed cells, denoted as
rat 2.2 cells, were generated by transfection of rat-1 fibroblasts
as described (see Seeburg, P. H., Colby, W. W., Capon, D. J.,
Goeddel, D. V., and Levinson, A. D. Nature 312, 71 (1984)). Cells
that overgrew the monolayer were extracted and plated onto agar to
obtain a cell line displaying a fully transformed phenotype.
Untransformed rat-1 fibroblasts and src-transformed rat-1
fibroblasts were also treated. The src-transformed cells were
generated by co-transfection of rat-1 fibroblasts with pSV3.
Neo.src, a vector containing the gene for src as well as the gene
that confers G418 resistance, both under control of SV40 early
promoters. G418-resistant cell clones that diplayed a transformed
phenotype were used to generate the cell line. On day 0, cells were
plated in monolayer culture at 3.times.103 cells per well (24-well
plates) in 1 ml DMEM supplemented with 10% FCS, 100 U/ml
penicillin, 10 .mu.g/ml streptomycin, 2% DMSO, and 0.5 mM DTT with
and without 200 .mu.M inhibitor. On day 3, the cells were refed
with the same medium. On day 5, the cells were photographed under
contrast at a magnification of 100.times.. Incubation of ras
transformed rat 2.2 cells with compound 27B (denoted as BZA-2B in
the figure) for 5 days reversed the transformed phenotype (FIG. 6B)
while compound 27A (denoted as BZA-2A in the figure) had no effect
(FIG. 6A). Clearly, after 5 days, the rat 2.2 cells display a more
normal, flattened cell morphology and grew to lower cell density.
This change in morphology is remarkably similar to that seen
following injection of anti-ras antibodies into ras-transformed
cells (see Feramisco, J. R. et al Nature 314, 639 (1985)). Rat-1
fibroblasts transformed with the src oncogene also grew in a
multilayered pattern (FIG. 6C), but this was not affected by
compound 27A or compound 27B (FIG. 6D). The compounds also had no
apparent effect on the morphology of untransformed rat-1
fibroblasts (FIG. 6E and F).
EXAMPLE 57
Effect of Farnesyltransferase inhibition on cell growth in
transformed and untransformed cells
To examine the effects of farnesyltransferase inhibition on cell
growth, three cell lines (ras-transformed rat-1, src-transformed
rat-1, and parental rat-1 cells) were seeded (see FIG. 7) at low
density and allowed to grow for 10 days in the absence or presence
of increasing concentrations of BZA-5B (compound 37). See Example
56 for description of cells. On day 0, cells were plated in
monolayer culture at 2.times.103 cells per well (24-well plates) in
1 ml DMEM supplemented with 10% fetal calf serum, 100 U/ml
penicillin, 100 .mu.g/ml streptomycin, 0.5 mM DTT, 0.025% DMSO, and
the indicated concentration of BZA-5B (compound 37). On days 3 and
7, cells were washed and refed with the same medium. At the time
indicated in FIG. 7, cells were harvested by trypsinization and
counted in a Coulter counter. Panels A, C, E show the growth rate
of each cell line in the absence of inhibitor. Panels B, D, F show
the inhibition of growth as a function of concentration of BZA-5B
at each time point. The "100% values" correspond to the appropriate
cell numbers in Panels A, C, E at the indicated time. Each value
represents a single incubation. As shown in the panels A, C, and E,
all three cell lines grew logarithmically with no added inhibitor.
In the presence of BZA-5B, the growth of ras-transformed
fibroblasts was inhibited in a time- and dosage-dependent manner,
reaching .about.90% inhibition after 10 days in the presence of 25
.mu.M BZA-5B (Panel B). The growth of src-transformed (Panel D) and
untransformed cells (Panel F) were not affected at concentrations
of BZA-5B up to 25 .mu.M. These results show that this inhibitor
specifically slows the growth of intact ras-transformed cells with
no effect on "normal" non-transformed cells, suggesting that these
inhibitors are potential treatments for tumors in which oncogenic
ras may play a role.
All references cited herein are expressly incorporated by
reference.
* * * * *